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anti kdm6a d3q1i rabbit antibody  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc anti kdm6a d3q1i rabbit antibody
    (A) Immunoprecipitation followed by western blotting analysis showing ETV2 and BAF155 association with BRG1, UTX ( <t>Kdm6a</t> ), and EP300. (B) Analysis of PDGFRα and FLK1 expression in day 4 wild-type and Baf155-KO EBs. The frequencies of DP and SP generation are quantified and shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (C) ETV2 ChIP sequencing peaks (highlighted) nearest the candidate target hemangiogenic genes. (D) ChIP-qPCR analysis showing BAF155 binding at ETV2 target loci. (E) ChIP-qPCR analysis showing ETV2 binding at target loci in wild-type and Baf155-KO EB cells. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (F) Expression of ETV2-regulated genes in wild-type and Baf155 -KO EB cells determined by RT-qPCR and normalized to β-actin housekeeping gene expression. The data are represented as the average ± standard deviation. The significance of data was determined using one-way ANOVA. (G) Hematopoietic colony-forming assays performed using day 4 wild-type and Baf155 EBs. The number of hematopoietic colonies formed is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (H) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Flk -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (I) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Etv2 -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. Significance of data: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ns, non-significant.
    Anti Kdm6a D3q1i Rabbit Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/kdm6a+antibody/UTX+Rabbit+mAb/pmc13003410-122-0-5
    Average 95 stars, based on 126 article reviews
    anti kdm6a d3q1i rabbit antibody - by Bioz Stars, 2026-09
    95/100 stars

    Images

    1) Product Images from "Distinct contributions of Etv2 + and Flk1 + progenitors to endothelial, hematopoietic, and cardiac lineages"

    Article Title: Distinct contributions of Etv2 + and Flk1 + progenitors to endothelial, hematopoietic, and cardiac lineages

    Journal: Cell reports

    doi: 10.1016/j.celrep.2025.116733

    (A) Immunoprecipitation followed by western blotting analysis showing ETV2 and BAF155 association with BRG1, UTX ( Kdm6a ), and EP300. (B) Analysis of PDGFRα and FLK1 expression in day 4 wild-type and Baf155-KO EBs. The frequencies of DP and SP generation are quantified and shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (C) ETV2 ChIP sequencing peaks (highlighted) nearest the candidate target hemangiogenic genes. (D) ChIP-qPCR analysis showing BAF155 binding at ETV2 target loci. (E) ChIP-qPCR analysis showing ETV2 binding at target loci in wild-type and Baf155-KO EB cells. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (F) Expression of ETV2-regulated genes in wild-type and Baf155 -KO EB cells determined by RT-qPCR and normalized to β-actin housekeeping gene expression. The data are represented as the average ± standard deviation. The significance of data was determined using one-way ANOVA. (G) Hematopoietic colony-forming assays performed using day 4 wild-type and Baf155 EBs. The number of hematopoietic colonies formed is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (H) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Flk -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (I) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Etv2 -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. Significance of data: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ns, non-significant.
    Figure Legend Snippet: (A) Immunoprecipitation followed by western blotting analysis showing ETV2 and BAF155 association with BRG1, UTX ( Kdm6a ), and EP300. (B) Analysis of PDGFRα and FLK1 expression in day 4 wild-type and Baf155-KO EBs. The frequencies of DP and SP generation are quantified and shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (C) ETV2 ChIP sequencing peaks (highlighted) nearest the candidate target hemangiogenic genes. (D) ChIP-qPCR analysis showing BAF155 binding at ETV2 target loci. (E) ChIP-qPCR analysis showing ETV2 binding at target loci in wild-type and Baf155-KO EB cells. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (F) Expression of ETV2-regulated genes in wild-type and Baf155 -KO EB cells determined by RT-qPCR and normalized to β-actin housekeeping gene expression. The data are represented as the average ± standard deviation. The significance of data was determined using one-way ANOVA. (G) Hematopoietic colony-forming assays performed using day 4 wild-type and Baf155 EBs. The number of hematopoietic colonies formed is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (H) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Flk -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (I) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Etv2 -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. Significance of data: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ns, non-significant.

    Techniques Used: Immunoprecipitation, Western Blot, Expressing, Standard Deviation, ChIP-sequencing, ChIP-qPCR, Binding Assay, Quantitative RT-PCR, Gene Expression, Control, Generated

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    Biomarker Discovery:

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    Knock-Out:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
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    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
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    Knockdown:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
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    Mutagenesis:

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    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
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    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Membrane:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Staining:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Fluorescence:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Chromatin Immunoprecipitation:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Flow Cytometry:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Magnetic Resonance Imaging:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Functional Assay:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Immunoprecipitation:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Expressing:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Western Blot:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Transfection:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Cell Surface Receptor Assay:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    ChIP-qPCR:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Quantitative RT-PCR:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Residue:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    shRNA:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Infection:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Control:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Luciferase:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).

    Synthesized:

    Article Title: Regulation of Epigenetic Modifiers, Including KDM6B, by Interferon-γ and Interleukin-4 in Human Macrophages
    Article Snippet: The antibodies used were Phospho-Rb-Ser807/811, E2F1, KDM6A, and SMYD3 (Cell Signaling), Histone H3 and Histone H3-Phospho S10 (Abcam), GAPDH (Millipore), and p27 kip1 (BD Biosciences).

    Article Title: Epigenetic regulation of CD38/CD48 by KDM6A mediates NK cell response in multiple myeloma
    Article Snippet: If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. ary April 2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Antibodies were obtained as follows: KDM6A (no. 33510, Cell Signaling Technology, 1:1000), CD38 (no. 51000, Cell Signaling Technology, 1:1000), CD48 (no. 29499, Cell Signaling Technology, 1:1000), GAPDH (no. 5174, Cell Signaling Technology, 1:1000), H3K27me3 (no. 9733, Cell Signaling Technology, 1:1000), anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)-linked Ab (no. 7074, Cell Signaling Technology, 1:2000), FITC anti-human CD38 (no. 356610, Biolegend, 1:20), APC anti-human CD38 (no. 356606, Biolegend, 1:20), FITC mouse IgG1 (no. 400110, Biolegend, 1:20), APC mouse IgG1 (no. 981806, Biolegend, 1:20), APC antihuman CD138 (no. 356506, Biolegend, 1:20), FITC anti-human IFN- (no. 502506, Biolegend, 1:20), APC anti-human CD48 (no. 336714, Biolegend, 1:20), Alexa Fluor 488 anti-human MICA/B (no. 320912, Biolegend, 1:20), APC anti-human CD56 (no. 985906, Biolegend, 1:20), APC anti-human HLA-A,B,C (no. 311409, Biolegend, 1:20), APC anti-human CD253 (no. 308209, Biolegend, 1:20), FITC anti-human CD319 (no. 331817, Biolegend, 1:20), FITC anti-human CD155 (no. 337627, Biolegend, 1:20), PE anti-human ULBP-2/5/6 (no. FAB1298P, R&D systems, 1:20), BV421 anti-human CD178 (no. 306411, Biolegend, 1:20).

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes
    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Article Title: Therapeutic targeting of histone lysine demethylase KDM4B blocks the growth of castration-resistant prostate cancer.
    Article Snippet: Following antibodies were used: pmTOR, mTOR, pS6, S6 kinase, KDM4B, Histone 3, cleaved PARP1, KDM6A, and KDM6B (Cell signaling), H3K9me3 and H3K27me3 (Abcam), GAPDH and SNAI2 (Santa Cruz), E-cadherin (BD company), SYP, PSA, and AR (cell Margue), and AR-V7 (Proteintech).



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    Inhibition of <t>KDM6A</t> prevents the H3K27 methyl-to-acetyl switch and gene activation induced by BRAF V600E inhibition. (A) Pharmacological inhibition of KDM6A by GSKJ4 in A375P cells increases H3K27me3 while reducing H3K27ac in a dose-dependent manner. Cells were treated with DMSO or GSKJ4 at the indicated concentration for 24 h. (B-C) GSKJ4 treatment inhibits the BRAF V600E inhibition–induced changes in H3K27me3 and H3K27ac in A375P cells, as shown by Western blot ( B ) and immunofluorescence ( C ). Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 24 h. (D) In A375P cells, KDM6A inhibition blunts the PLX4032-mediated alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters, as determined by ChIP. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. (E) GSKJ4 treatment abrogates the upregulation of EZH2 target genes (PGC1α, DCT, MET, WDR19) by BRAF V600E inhibition in A375P cells. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. ( F ) KDM6A knockout efficiency in A375P cells was confirmed by qRT-PCR and Western blot. (G-I) CRISPR-mediated KDM6A deletion prevents the changes in H3K27me3 and H3K27ac triggered by BRAF V600E inhibition in A375P cells, as shown by Western blot ( G ) and immunofluorescence ( H and I ). Cells were treated with DMSO or 2 μM PLX4032 for 24 h. (J) KDM6A deletion abolishes the alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. (K) CRISPR-mediated KDM6A deletion suppresses the BRAF V600E inhibition–induced upregulation of EZH2 target genes in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. Data are presented as mean ± SEM and representative of two to three independent experiments with similar results ( n = 3/group/experiment). * P < 0.05, ** P < 0.01, *** P < 0.001 by Student’s t-test in (F) and by two-way ANOVA in (C), (D), (E), (I), (J), and (K).
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    Inhibition of <t>KDM6A</t> prevents the H3K27 methyl-to-acetyl switch and gene activation induced by BRAF V600E inhibition. (A) Pharmacological inhibition of KDM6A by GSKJ4 in A375P cells increases H3K27me3 while reducing H3K27ac in a dose-dependent manner. Cells were treated with DMSO or GSKJ4 at the indicated concentration for 24 h. (B-C) GSKJ4 treatment inhibits the BRAF V600E inhibition–induced changes in H3K27me3 and H3K27ac in A375P cells, as shown by Western blot ( B ) and immunofluorescence ( C ). Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 24 h. (D) In A375P cells, KDM6A inhibition blunts the PLX4032-mediated alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters, as determined by ChIP. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. (E) GSKJ4 treatment abrogates the upregulation of EZH2 target genes (PGC1α, DCT, MET, WDR19) by BRAF V600E inhibition in A375P cells. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. ( F ) KDM6A knockout efficiency in A375P cells was confirmed by qRT-PCR and Western blot. (G-I) CRISPR-mediated KDM6A deletion prevents the changes in H3K27me3 and H3K27ac triggered by BRAF V600E inhibition in A375P cells, as shown by Western blot ( G ) and immunofluorescence ( H and I ). Cells were treated with DMSO or 2 μM PLX4032 for 24 h. (J) KDM6A deletion abolishes the alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. (K) CRISPR-mediated KDM6A deletion suppresses the BRAF V600E inhibition–induced upregulation of EZH2 target genes in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. Data are presented as mean ± SEM and representative of two to three independent experiments with similar results ( n = 3/group/experiment). * P < 0.05, ** P < 0.01, *** P < 0.001 by Student’s t-test in (F) and by two-way ANOVA in (C), (D), (E), (I), (J), and (K).
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    Inhibition of <t>KDM6A</t> prevents the H3K27 methyl-to-acetyl switch and gene activation induced by BRAF V600E inhibition. (A) Pharmacological inhibition of KDM6A by GSKJ4 in A375P cells increases H3K27me3 while reducing H3K27ac in a dose-dependent manner. Cells were treated with DMSO or GSKJ4 at the indicated concentration for 24 h. (B-C) GSKJ4 treatment inhibits the BRAF V600E inhibition–induced changes in H3K27me3 and H3K27ac in A375P cells, as shown by Western blot ( B ) and immunofluorescence ( C ). Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 24 h. (D) In A375P cells, KDM6A inhibition blunts the PLX4032-mediated alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters, as determined by ChIP. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. (E) GSKJ4 treatment abrogates the upregulation of EZH2 target genes (PGC1α, DCT, MET, WDR19) by BRAF V600E inhibition in A375P cells. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. ( F ) KDM6A knockout efficiency in A375P cells was confirmed by qRT-PCR and Western blot. (G-I) CRISPR-mediated KDM6A deletion prevents the changes in H3K27me3 and H3K27ac triggered by BRAF V600E inhibition in A375P cells, as shown by Western blot ( G ) and immunofluorescence ( H and I ). Cells were treated with DMSO or 2 μM PLX4032 for 24 h. (J) KDM6A deletion abolishes the alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. (K) CRISPR-mediated KDM6A deletion suppresses the BRAF V600E inhibition–induced upregulation of EZH2 target genes in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. Data are presented as mean ± SEM and representative of two to three independent experiments with similar results ( n = 3/group/experiment). * P < 0.05, ** P < 0.01, *** P < 0.001 by Student’s t-test in (F) and by two-way ANOVA in (C), (D), (E), (I), (J), and (K).
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    <t>kdm6a</t> is required for HSPC emergence in zebrafish. A,B) WISH for runx1 A) and %phenotype distribution B) in embryos at 28 hpf as indicated (n = 4, mean ± SD, Student's t test). C,D) WISH for runx1 C) and %phenotype distribution D) in embryos at 30 hpf as indicated (n = 4, mean ± SD, Student's t test). E,F) WISH for cmyb E) and %phenotype distribution F) in embryos at 30 hpf as indicated ( n = 4, mean ± SD, Student's t test). G,H) WISH for cmyb G) and %phenotype distribution H) in embryos at 3 dpf as indicated (n = 4, mean ± SD, Student's t test). I,J) WISH for rag1 I) and %phenotype distribution J) in embryos at 4 dpf as indicated (n = 4, mean ± SD, Student's t test). K) Flow cytometry plots of kdrl :mCherry + ; runx1 :eGFP + double positive cells at 28 hpf (left). Graphs depicting the percentage of kdrl :mCherry + ; runx1 :eGFP + hemogenic endothelial cells per embryo at 28 hpf (right) ( n = 4, mean ± SD, Student's t test). L) Confocal imaging of kdrl :mCherry + ; cmyb :eGFP + hemogenic endothelial cells in AGM at 48 hpf (left, white arrowheads). Graphs depicting the number of kdrl :mCherry + ; cmyb :eGFP + cells per embryo at 48 hpf (right) ( n = 5, mean ± SD, Student's t test). M) Flow cytometry plots of cd41 :eGFP low cells at 3 dpf (left). Graphs depicting the percentage of cd41 :eGFP low HSPCs per embryo at 3 dpf (right) ( n = 5, mean ± SD, Student's t test). N,O) WISH for runx1 N) and %phenotype distribution O) in embryos at 28 hpf as indicated ( n = 4, mean ± SD, one‐way ANOVA). ** p <0.01; *** p < 0.001.
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    <t>Kdm6a</t> KO results in Xist downregulation. A Principal component analysis (PCA) based on expression of autosomal or X-linked genes in two Tsix -stop wt clones and two Kdm6a KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) before (D0) and after differentiation (D15). Only genes with a minimum of 2.5 CPM in 4 libraries were considered. PCA shows separation of clones based on Kdm6a KO status mainly after differentiation. PCA figures were generated by iDEP.95. B Plots of diploid gene expression from a representative autosome (Chr1, left) with similar gene content as the X, and from the X chromosome (right) in Tsix -stop wt and KO clones (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D15. Kdm6a and the XIC are highlighted. Log 2 TPM fold change (KO/wt) is shown. Genes with a decrease of expression in KO versus wt are in blue, and with an increase in red. C qRT-PCR of average Xist expression normalized to Actinβ during ES differentiation in three Tsix -stop wt samples, two Kdm6a ΔEΔE KO clones, and one Kdm6a ΔwtΔE clone. p-value is derived from comparing wt to Kdm6a ΔEΔE biological replicates. Divergence of Xist expression between wt and KO cells begins at D2 and is largest at D7 (p = 0.04). The Kdm6a wt/ΔE4 heterozygous KO clone behaves similarly to the homozygous clones. D qRT-PCR of Xist expression in E8 wt and KO clone (E8- Kdm6a ΔPΔP13 ). Expression is normalized to Actinβ . Xist is significantly lower at D2 and D4 in KO cells. P-values derived from two technical replicates of wt and KO cells (**p < 0.01;*p < 0.05). E RNA-FISH with a probe specific for Xist RNA labeled in green in Tsix -stop wt and KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ). Examples of nuclei are shown together with histograms for quantification of signal type (pinpoint, no signal or cloud). Xist clouds are present in 74% of D15 wt cells, but significantly decrease in KO clones (cloud compared to no signal; ***p < 0.00001 for T six-Kdm6a ΔEΔE17 and < 0.0001 for Tsix-Kdm6a ΔEΔE21 fisher’s exact test), while the frequency of pinpoint Xist signals increases in KO clones (pinpoint in wt compared to pinpoint in KO; ***p < 0.00001 for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 fisher’s exact test). Nuclei are counterstained with Hoechst 33,342. Scale bar = 10 µm
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    <t>Kdm6a</t> KO results in Xist downregulation. A Principal component analysis (PCA) based on expression of autosomal or X-linked genes in two Tsix -stop wt clones and two Kdm6a KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) before (D0) and after differentiation (D15). Only genes with a minimum of 2.5 CPM in 4 libraries were considered. PCA shows separation of clones based on Kdm6a KO status mainly after differentiation. PCA figures were generated by iDEP.95. B Plots of diploid gene expression from a representative autosome (Chr1, left) with similar gene content as the X, and from the X chromosome (right) in Tsix -stop wt and KO clones (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D15. Kdm6a and the XIC are highlighted. Log 2 TPM fold change (KO/wt) is shown. Genes with a decrease of expression in KO versus wt are in blue, and with an increase in red. C qRT-PCR of average Xist expression normalized to Actinβ during ES differentiation in three Tsix -stop wt samples, two Kdm6a ΔEΔE KO clones, and one Kdm6a ΔwtΔE clone. p-value is derived from comparing wt to Kdm6a ΔEΔE biological replicates. Divergence of Xist expression between wt and KO cells begins at D2 and is largest at D7 (p = 0.04). The Kdm6a wt/ΔE4 heterozygous KO clone behaves similarly to the homozygous clones. D qRT-PCR of Xist expression in E8 wt and KO clone (E8- Kdm6a ΔPΔP13 ). Expression is normalized to Actinβ . Xist is significantly lower at D2 and D4 in KO cells. P-values derived from two technical replicates of wt and KO cells (**p < 0.01;*p < 0.05). E RNA-FISH with a probe specific for Xist RNA labeled in green in Tsix -stop wt and KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ). Examples of nuclei are shown together with histograms for quantification of signal type (pinpoint, no signal or cloud). Xist clouds are present in 74% of D15 wt cells, but significantly decrease in KO clones (cloud compared to no signal; ***p < 0.00001 for T six-Kdm6a ΔEΔE17 and < 0.0001 for Tsix-Kdm6a ΔEΔE21 fisher’s exact test), while the frequency of pinpoint Xist signals increases in KO clones (pinpoint in wt compared to pinpoint in KO; ***p < 0.00001 for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 fisher’s exact test). Nuclei are counterstained with Hoechst 33,342. Scale bar = 10 µm
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    <t>Kdm6a</t> KO results in Xist downregulation. A Principal component analysis (PCA) based on expression of autosomal or X-linked genes in two Tsix -stop wt clones and two Kdm6a KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) before (D0) and after differentiation (D15). Only genes with a minimum of 2.5 CPM in 4 libraries were considered. PCA shows separation of clones based on Kdm6a KO status mainly after differentiation. PCA figures were generated by iDEP.95. B Plots of diploid gene expression from a representative autosome (Chr1, left) with similar gene content as the X, and from the X chromosome (right) in Tsix -stop wt and KO clones (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D15. Kdm6a and the XIC are highlighted. Log 2 TPM fold change (KO/wt) is shown. Genes with a decrease of expression in KO versus wt are in blue, and with an increase in red. C qRT-PCR of average Xist expression normalized to Actinβ during ES differentiation in three Tsix -stop wt samples, two Kdm6a ΔEΔE KO clones, and one Kdm6a ΔwtΔE clone. p-value is derived from comparing wt to Kdm6a ΔEΔE biological replicates. Divergence of Xist expression between wt and KO cells begins at D2 and is largest at D7 (p = 0.04). The Kdm6a wt/ΔE4 heterozygous KO clone behaves similarly to the homozygous clones. D qRT-PCR of Xist expression in E8 wt and KO clone (E8- Kdm6a ΔPΔP13 ). Expression is normalized to Actinβ . Xist is significantly lower at D2 and D4 in KO cells. P-values derived from two technical replicates of wt and KO cells (**p < 0.01;*p < 0.05). E RNA-FISH with a probe specific for Xist RNA labeled in green in Tsix -stop wt and KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ). Examples of nuclei are shown together with histograms for quantification of signal type (pinpoint, no signal or cloud). Xist clouds are present in 74% of D15 wt cells, but significantly decrease in KO clones (cloud compared to no signal; ***p < 0.00001 for T six-Kdm6a ΔEΔE17 and < 0.0001 for Tsix-Kdm6a ΔEΔE21 fisher’s exact test), while the frequency of pinpoint Xist signals increases in KO clones (pinpoint in wt compared to pinpoint in KO; ***p < 0.00001 for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 fisher’s exact test). Nuclei are counterstained with Hoechst 33,342. Scale bar = 10 µm
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    Image Search Results


    (A) Immunoprecipitation followed by western blotting analysis showing ETV2 and BAF155 association with BRG1, UTX ( Kdm6a ), and EP300. (B) Analysis of PDGFRα and FLK1 expression in day 4 wild-type and Baf155-KO EBs. The frequencies of DP and SP generation are quantified and shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (C) ETV2 ChIP sequencing peaks (highlighted) nearest the candidate target hemangiogenic genes. (D) ChIP-qPCR analysis showing BAF155 binding at ETV2 target loci. (E) ChIP-qPCR analysis showing ETV2 binding at target loci in wild-type and Baf155-KO EB cells. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (F) Expression of ETV2-regulated genes in wild-type and Baf155 -KO EB cells determined by RT-qPCR and normalized to β-actin housekeeping gene expression. The data are represented as the average ± standard deviation. The significance of data was determined using one-way ANOVA. (G) Hematopoietic colony-forming assays performed using day 4 wild-type and Baf155 EBs. The number of hematopoietic colonies formed is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (H) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Flk -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (I) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Etv2 -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. Significance of data: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ns, non-significant.

    Journal: Cell reports

    Article Title: Distinct contributions of Etv2 + and Flk1 + progenitors to endothelial, hematopoietic, and cardiac lineages

    doi: 10.1016/j.celrep.2025.116733

    Figure Lengend Snippet: (A) Immunoprecipitation followed by western blotting analysis showing ETV2 and BAF155 association with BRG1, UTX ( Kdm6a ), and EP300. (B) Analysis of PDGFRα and FLK1 expression in day 4 wild-type and Baf155-KO EBs. The frequencies of DP and SP generation are quantified and shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (C) ETV2 ChIP sequencing peaks (highlighted) nearest the candidate target hemangiogenic genes. (D) ChIP-qPCR analysis showing BAF155 binding at ETV2 target loci. (E) ChIP-qPCR analysis showing ETV2 binding at target loci in wild-type and Baf155-KO EB cells. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (F) Expression of ETV2-regulated genes in wild-type and Baf155 -KO EB cells determined by RT-qPCR and normalized to β-actin housekeeping gene expression. The data are represented as the average ± standard deviation. The significance of data was determined using one-way ANOVA. (G) Hematopoietic colony-forming assays performed using day 4 wild-type and Baf155 EBs. The number of hematopoietic colonies formed is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (H) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Flk -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. (I) Hematopoietic colony-forming assays performed using ~E8.5 Baf155 Δ Etv2 -cKO and control embryos. The number of BFU-E (erythroid) and myeloid colonies generated is shown. The data are represented as the average ± standard deviation. The significance of data was determined using an unpaired t test. Significance of data: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ns, non-significant.

    Article Snippet: anti-KDM6A [D3Q1I] rabbit antibody , Cell Signaling Technologies , Cat# 33510; RRID: AB_2721244.

    Techniques: Immunoprecipitation, Western Blot, Expressing, Standard Deviation, ChIP-sequencing, ChIP-qPCR, Binding Assay, Quantitative RT-PCR, Gene Expression, Control, Generated

    Inhibition of KDM6A prevents the H3K27 methyl-to-acetyl switch and gene activation induced by BRAF V600E inhibition. (A) Pharmacological inhibition of KDM6A by GSKJ4 in A375P cells increases H3K27me3 while reducing H3K27ac in a dose-dependent manner. Cells were treated with DMSO or GSKJ4 at the indicated concentration for 24 h. (B-C) GSKJ4 treatment inhibits the BRAF V600E inhibition–induced changes in H3K27me3 and H3K27ac in A375P cells, as shown by Western blot ( B ) and immunofluorescence ( C ). Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 24 h. (D) In A375P cells, KDM6A inhibition blunts the PLX4032-mediated alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters, as determined by ChIP. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. (E) GSKJ4 treatment abrogates the upregulation of EZH2 target genes (PGC1α, DCT, MET, WDR19) by BRAF V600E inhibition in A375P cells. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. ( F ) KDM6A knockout efficiency in A375P cells was confirmed by qRT-PCR and Western blot. (G-I) CRISPR-mediated KDM6A deletion prevents the changes in H3K27me3 and H3K27ac triggered by BRAF V600E inhibition in A375P cells, as shown by Western blot ( G ) and immunofluorescence ( H and I ). Cells were treated with DMSO or 2 μM PLX4032 for 24 h. (J) KDM6A deletion abolishes the alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. (K) CRISPR-mediated KDM6A deletion suppresses the BRAF V600E inhibition–induced upregulation of EZH2 target genes in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. Data are presented as mean ± SEM and representative of two to three independent experiments with similar results ( n = 3/group/experiment). * P < 0.05, ** P < 0.01, *** P < 0.001 by Student’s t-test in (F) and by two-way ANOVA in (C), (D), (E), (I), (J), and (K).

    Journal: Neoplasia (New York, N.Y.)

    Article Title: A methyl-to-acetyl switch in H3K27 drives metabolic reprogramming and resistance to BRAF V600E inhibition in melanoma

    doi: 10.1016/j.neo.2025.101223

    Figure Lengend Snippet: Inhibition of KDM6A prevents the H3K27 methyl-to-acetyl switch and gene activation induced by BRAF V600E inhibition. (A) Pharmacological inhibition of KDM6A by GSKJ4 in A375P cells increases H3K27me3 while reducing H3K27ac in a dose-dependent manner. Cells were treated with DMSO or GSKJ4 at the indicated concentration for 24 h. (B-C) GSKJ4 treatment inhibits the BRAF V600E inhibition–induced changes in H3K27me3 and H3K27ac in A375P cells, as shown by Western blot ( B ) and immunofluorescence ( C ). Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 24 h. (D) In A375P cells, KDM6A inhibition blunts the PLX4032-mediated alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters, as determined by ChIP. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. (E) GSKJ4 treatment abrogates the upregulation of EZH2 target genes (PGC1α, DCT, MET, WDR19) by BRAF V600E inhibition in A375P cells. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. ( F ) KDM6A knockout efficiency in A375P cells was confirmed by qRT-PCR and Western blot. (G-I) CRISPR-mediated KDM6A deletion prevents the changes in H3K27me3 and H3K27ac triggered by BRAF V600E inhibition in A375P cells, as shown by Western blot ( G ) and immunofluorescence ( H and I ). Cells were treated with DMSO or 2 μM PLX4032 for 24 h. (J) KDM6A deletion abolishes the alterations in H3K27ac occupancy at the PGC1α, DCT, MET, and WDR19 promoters in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. (K) CRISPR-mediated KDM6A deletion suppresses the BRAF V600E inhibition–induced upregulation of EZH2 target genes in A375P cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. Data are presented as mean ± SEM and representative of two to three independent experiments with similar results ( n = 3/group/experiment). * P < 0.05, ** P < 0.01, *** P < 0.001 by Student’s t-test in (F) and by two-way ANOVA in (C), (D), (E), (I), (J), and (K).

    Article Snippet: Antibodies included: KDM6A (NOVUS, #NBP1-80628), BRD4 (HUABIO, #HA722785), HRP-conjugated β-Actin (ABclonal, #AC043), EZH2 (D2C9) (Cell Signaling Technology, #5246S), Histone H3 (D1H2) (CST, #4499S), H3K27me3 (C36B11) (CST, #9733S), H3K27ac (D5E4) (CST, #8173S), Cleaved PARP (HUABIO, #ET1608-10), Alpha Tubulin (HUABIO, # ER130905 ), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) (CST, #4370S), and p44/42 MAPK (Erk1/2) (137F5) (CST, #4695).

    Techniques: Inhibition, Activation Assay, Concentration Assay, Western Blot, Immunofluorescence, Knock-Out, Quantitative RT-PCR, CRISPR

    Blockade of the KDM6A–H3K27ac–BRD4 axis reprograms melanoma metabolism. (A) Pharmacological inhibition of BRD4 by JQ1 attenuates the PLX4032-induced mitochondrial metabolic program in A375P cells. Cells were treated with DMSO, 2 μM JQ1, 2 μM PLX4032, or both for 16 h. (B) CRISPR-mediated BRD4 depletion suppresses the mitochondrial metabolic program triggered by PLX4032 in A375P melanoma cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. (C-D) BRD4 inactivation reduces mitochondrial content ( C ) and respiration ( D ) induced by BRAF V600E inhibition in A375P cells, as measured by MitoTracker staining and a Clark-type oxygen electrode, respectively. Cells were treated with DMSO, 2 μM JQ1, 2 μM PLX4032, or both for 24 h. (E) Pharmacological inhibition of KDM6A activity by GSKJ4 prevents the PLX4032-mediated increase in the mitochondrial metabolic program in A375P cells. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. (F) CRISPR-mediated KDM6A depletion suppresses the PLX4032-induced mitochondrial metabolic program in A375P melanoma cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. (G-H) KDM6A inhibition decreases mitochondrial content ( G ) and respiration ( H ) following BRAF V600E inhibition in A375P cells. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 24 h. Data are represented as mean ± SEM, and representative of two to three independent experiments with similar results ( n = 3/group/repeat). * P < 0.05, ** P < 0.01, and *** P < 0.001 by two-way ANOVA.

    Journal: Neoplasia (New York, N.Y.)

    Article Title: A methyl-to-acetyl switch in H3K27 drives metabolic reprogramming and resistance to BRAF V600E inhibition in melanoma

    doi: 10.1016/j.neo.2025.101223

    Figure Lengend Snippet: Blockade of the KDM6A–H3K27ac–BRD4 axis reprograms melanoma metabolism. (A) Pharmacological inhibition of BRD4 by JQ1 attenuates the PLX4032-induced mitochondrial metabolic program in A375P cells. Cells were treated with DMSO, 2 μM JQ1, 2 μM PLX4032, or both for 16 h. (B) CRISPR-mediated BRD4 depletion suppresses the mitochondrial metabolic program triggered by PLX4032 in A375P melanoma cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. (C-D) BRD4 inactivation reduces mitochondrial content ( C ) and respiration ( D ) induced by BRAF V600E inhibition in A375P cells, as measured by MitoTracker staining and a Clark-type oxygen electrode, respectively. Cells were treated with DMSO, 2 μM JQ1, 2 μM PLX4032, or both for 24 h. (E) Pharmacological inhibition of KDM6A activity by GSKJ4 prevents the PLX4032-mediated increase in the mitochondrial metabolic program in A375P cells. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 16 h. (F) CRISPR-mediated KDM6A depletion suppresses the PLX4032-induced mitochondrial metabolic program in A375P melanoma cells. Cells were treated with DMSO or 2 μM PLX4032 for 16 h. (G-H) KDM6A inhibition decreases mitochondrial content ( G ) and respiration ( H ) following BRAF V600E inhibition in A375P cells. Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 24 h. Data are represented as mean ± SEM, and representative of two to three independent experiments with similar results ( n = 3/group/repeat). * P < 0.05, ** P < 0.01, and *** P < 0.001 by two-way ANOVA.

    Article Snippet: Antibodies included: KDM6A (NOVUS, #NBP1-80628), BRD4 (HUABIO, #HA722785), HRP-conjugated β-Actin (ABclonal, #AC043), EZH2 (D2C9) (Cell Signaling Technology, #5246S), Histone H3 (D1H2) (CST, #4499S), H3K27me3 (C36B11) (CST, #9733S), H3K27ac (D5E4) (CST, #8173S), Cleaved PARP (HUABIO, #ET1608-10), Alpha Tubulin (HUABIO, # ER130905 ), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) (CST, #4370S), and p44/42 MAPK (Erk1/2) (137F5) (CST, #4695).

    Techniques: Inhibition, CRISPR, Staining, Activity Assay

    The H3K27 methyl-to-acetyl switch enables melanoma to survive BRAF V600E inhibition. (A) Basal expression levels of BRD4 or KDM6s negatively correlate with BRAF V600E inhibitor sensitivity across a panel of human melanoma cell lines. (B) In melanoma patients, basal BRD4 and KDM6s expression in pre-treatment samples negatively correlates with tumor response to BRAF V600E /MEK inhibitors. (C-D) CRISPR-mediated BRD4 depletion reduces melanoma cell survival ( C ) by increasing apoptotic cell death ( D ) following BRAF V600E inhibition. Cells were treated with DMSO or 2 μM PLX4032 for 24 h. (E-F) CRISPR-mediated KDM6A depletion similarly decreases melanoma survival ( E ) and enhances apoptotic cell death ( F ) in response to BRAF V600E inhibition. Cells were treated with DMSO or 2 μM PLX4032 for 24 h. (G) CRISPR-mediated KDM6A depletion in BRAF V600E -mutant YUMM1.7 melanoma cells significantly improves the in vivo response to BRAF V600E inhibition. The diagram illustrates the treatment strategy. The following group sizes were used: sgSCR_Vehicle ( n = 7), sgSCR_PLX4032 ( n = 8), sgKdm6a_Vehicle ( n = 6), and sgKdm6a_PLX4032 ( n = 9). In (A) and (B), each data point represents individual samples. In all other panels, data are presented as mean ± SEM, and are representative of two to three independent experiments with similar results in (C) and (E) ( n = 3/group/repeat). Statistical significance was determined by Pearson correlation in (A) and (B), or by two-way ANOVA with * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Journal: Neoplasia (New York, N.Y.)

    Article Title: A methyl-to-acetyl switch in H3K27 drives metabolic reprogramming and resistance to BRAF V600E inhibition in melanoma

    doi: 10.1016/j.neo.2025.101223

    Figure Lengend Snippet: The H3K27 methyl-to-acetyl switch enables melanoma to survive BRAF V600E inhibition. (A) Basal expression levels of BRD4 or KDM6s negatively correlate with BRAF V600E inhibitor sensitivity across a panel of human melanoma cell lines. (B) In melanoma patients, basal BRD4 and KDM6s expression in pre-treatment samples negatively correlates with tumor response to BRAF V600E /MEK inhibitors. (C-D) CRISPR-mediated BRD4 depletion reduces melanoma cell survival ( C ) by increasing apoptotic cell death ( D ) following BRAF V600E inhibition. Cells were treated with DMSO or 2 μM PLX4032 for 24 h. (E-F) CRISPR-mediated KDM6A depletion similarly decreases melanoma survival ( E ) and enhances apoptotic cell death ( F ) in response to BRAF V600E inhibition. Cells were treated with DMSO or 2 μM PLX4032 for 24 h. (G) CRISPR-mediated KDM6A depletion in BRAF V600E -mutant YUMM1.7 melanoma cells significantly improves the in vivo response to BRAF V600E inhibition. The diagram illustrates the treatment strategy. The following group sizes were used: sgSCR_Vehicle ( n = 7), sgSCR_PLX4032 ( n = 8), sgKdm6a_Vehicle ( n = 6), and sgKdm6a_PLX4032 ( n = 9). In (A) and (B), each data point represents individual samples. In all other panels, data are presented as mean ± SEM, and are representative of two to three independent experiments with similar results in (C) and (E) ( n = 3/group/repeat). Statistical significance was determined by Pearson correlation in (A) and (B), or by two-way ANOVA with * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Article Snippet: Antibodies included: KDM6A (NOVUS, #NBP1-80628), BRD4 (HUABIO, #HA722785), HRP-conjugated β-Actin (ABclonal, #AC043), EZH2 (D2C9) (Cell Signaling Technology, #5246S), Histone H3 (D1H2) (CST, #4499S), H3K27me3 (C36B11) (CST, #9733S), H3K27ac (D5E4) (CST, #8173S), Cleaved PARP (HUABIO, #ET1608-10), Alpha Tubulin (HUABIO, # ER130905 ), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) (CST, #4370S), and p44/42 MAPK (Erk1/2) (137F5) (CST, #4695).

    Techniques: Inhibition, Expressing, CRISPR, Mutagenesis, In Vivo

    Targeting the H3K27 methyl-to-acetyl switch sensitizes melanoma to BRAF V600E inhibition. (A-B) BRD4 inactivation by JQ1 enhances the efficacy of PLX4032 in suppressing melanoma cell survival, as assessed by cell proliferation ( A ) and apoptosis (indicated by cleaved PARP1) ( B ). Cells were treated with DMSO, 2 μM JQ1, 2 μM PLX4032, or both for 24 h. (C-D) Pharmacological inhibition of KDM6A with GSKJ4 synergizes with PLX4032 to further suppress melanoma cell survival ( C ) and induce apoptotic cell death ( D ). Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 24 h. (E) The BRD4 inhibitor JQ1 significantly sensitizes BRAF V600E -mutant YUMM1.7 melanoma to PLX4032 in vivo . Vehicle ( n = 6), PLX4032 ( n = 5), JQ1 ( n = 5), PLX4032+JQ1 ( n = 10). The diagram illustrates the treatment strategy. (F) Inhibition of KDM6A by GSKJ4 significantly sensitizes BRAF V600E -mutant YUMM1.7 melanoma to PLX4032 in vivo . Vehicle ( n = 4), PLX4032 ( n = 4), GSKJ4 ( n = 4), PLX4032+GSKJ4 ( n = 5). (G-H) Treatment with GSKJ4 in combination with BRAF V600E inhibition reduces tumor progression ( G ) and prolongs survival ( H ) in a genetically engineered melanoma mouse model (Tyr::Cre ERT2 ;Braf CA ;Pten lox/lox ). Mice were induced with 4-hydroxytamoxifen (4-OHT; n = 5 per group for G, n = 6 per group for H). (I ) Diagram summarizing the interplay between BRAF V600E and the H3K27 epigenetic program in melanoma. Data are represented as mean ± SEM, and representative of two to three independent experiments with similar results in (A) and (C) ( n = 3/group/repeat). Statistical significance was determined by two-way ANOVA. Survival curves were estimated using the Kaplan–Meier method and compared using the log-rank (Mantel–Cox) test. * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Journal: Neoplasia (New York, N.Y.)

    Article Title: A methyl-to-acetyl switch in H3K27 drives metabolic reprogramming and resistance to BRAF V600E inhibition in melanoma

    doi: 10.1016/j.neo.2025.101223

    Figure Lengend Snippet: Targeting the H3K27 methyl-to-acetyl switch sensitizes melanoma to BRAF V600E inhibition. (A-B) BRD4 inactivation by JQ1 enhances the efficacy of PLX4032 in suppressing melanoma cell survival, as assessed by cell proliferation ( A ) and apoptosis (indicated by cleaved PARP1) ( B ). Cells were treated with DMSO, 2 μM JQ1, 2 μM PLX4032, or both for 24 h. (C-D) Pharmacological inhibition of KDM6A with GSKJ4 synergizes with PLX4032 to further suppress melanoma cell survival ( C ) and induce apoptotic cell death ( D ). Cells were treated with DMSO, 5 μM GSKJ4, 2 μM PLX4032, or both for 24 h. (E) The BRD4 inhibitor JQ1 significantly sensitizes BRAF V600E -mutant YUMM1.7 melanoma to PLX4032 in vivo . Vehicle ( n = 6), PLX4032 ( n = 5), JQ1 ( n = 5), PLX4032+JQ1 ( n = 10). The diagram illustrates the treatment strategy. (F) Inhibition of KDM6A by GSKJ4 significantly sensitizes BRAF V600E -mutant YUMM1.7 melanoma to PLX4032 in vivo . Vehicle ( n = 4), PLX4032 ( n = 4), GSKJ4 ( n = 4), PLX4032+GSKJ4 ( n = 5). (G-H) Treatment with GSKJ4 in combination with BRAF V600E inhibition reduces tumor progression ( G ) and prolongs survival ( H ) in a genetically engineered melanoma mouse model (Tyr::Cre ERT2 ;Braf CA ;Pten lox/lox ). Mice were induced with 4-hydroxytamoxifen (4-OHT; n = 5 per group for G, n = 6 per group for H). (I ) Diagram summarizing the interplay between BRAF V600E and the H3K27 epigenetic program in melanoma. Data are represented as mean ± SEM, and representative of two to three independent experiments with similar results in (A) and (C) ( n = 3/group/repeat). Statistical significance was determined by two-way ANOVA. Survival curves were estimated using the Kaplan–Meier method and compared using the log-rank (Mantel–Cox) test. * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Article Snippet: Antibodies included: KDM6A (NOVUS, #NBP1-80628), BRD4 (HUABIO, #HA722785), HRP-conjugated β-Actin (ABclonal, #AC043), EZH2 (D2C9) (Cell Signaling Technology, #5246S), Histone H3 (D1H2) (CST, #4499S), H3K27me3 (C36B11) (CST, #9733S), H3K27ac (D5E4) (CST, #8173S), Cleaved PARP (HUABIO, #ET1608-10), Alpha Tubulin (HUABIO, # ER130905 ), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) (CST, #4370S), and p44/42 MAPK (Erk1/2) (137F5) (CST, #4695).

    Techniques: Inhibition, Mutagenesis, In Vivo

    kdm6a is required for HSPC emergence in zebrafish. A,B) WISH for runx1 A) and %phenotype distribution B) in embryos at 28 hpf as indicated (n = 4, mean ± SD, Student's t test). C,D) WISH for runx1 C) and %phenotype distribution D) in embryos at 30 hpf as indicated (n = 4, mean ± SD, Student's t test). E,F) WISH for cmyb E) and %phenotype distribution F) in embryos at 30 hpf as indicated ( n = 4, mean ± SD, Student's t test). G,H) WISH for cmyb G) and %phenotype distribution H) in embryos at 3 dpf as indicated (n = 4, mean ± SD, Student's t test). I,J) WISH for rag1 I) and %phenotype distribution J) in embryos at 4 dpf as indicated (n = 4, mean ± SD, Student's t test). K) Flow cytometry plots of kdrl :mCherry + ; runx1 :eGFP + double positive cells at 28 hpf (left). Graphs depicting the percentage of kdrl :mCherry + ; runx1 :eGFP + hemogenic endothelial cells per embryo at 28 hpf (right) ( n = 4, mean ± SD, Student's t test). L) Confocal imaging of kdrl :mCherry + ; cmyb :eGFP + hemogenic endothelial cells in AGM at 48 hpf (left, white arrowheads). Graphs depicting the number of kdrl :mCherry + ; cmyb :eGFP + cells per embryo at 48 hpf (right) ( n = 5, mean ± SD, Student's t test). M) Flow cytometry plots of cd41 :eGFP low cells at 3 dpf (left). Graphs depicting the percentage of cd41 :eGFP low HSPCs per embryo at 3 dpf (right) ( n = 5, mean ± SD, Student's t test). N,O) WISH for runx1 N) and %phenotype distribution O) in embryos at 28 hpf as indicated ( n = 4, mean ± SD, one‐way ANOVA). ** p <0.01; *** p < 0.001.

    Journal: Advanced Science

    Article Title: KDM6A Deficiency Induces Myeloid Bias and Promotes CMML‐Like Disease Through JAK/STAT3 Activation by Repressing SOCS3

    doi: 10.1002/advs.202413091

    Figure Lengend Snippet: kdm6a is required for HSPC emergence in zebrafish. A,B) WISH for runx1 A) and %phenotype distribution B) in embryos at 28 hpf as indicated (n = 4, mean ± SD, Student's t test). C,D) WISH for runx1 C) and %phenotype distribution D) in embryos at 30 hpf as indicated (n = 4, mean ± SD, Student's t test). E,F) WISH for cmyb E) and %phenotype distribution F) in embryos at 30 hpf as indicated ( n = 4, mean ± SD, Student's t test). G,H) WISH for cmyb G) and %phenotype distribution H) in embryos at 3 dpf as indicated (n = 4, mean ± SD, Student's t test). I,J) WISH for rag1 I) and %phenotype distribution J) in embryos at 4 dpf as indicated (n = 4, mean ± SD, Student's t test). K) Flow cytometry plots of kdrl :mCherry + ; runx1 :eGFP + double positive cells at 28 hpf (left). Graphs depicting the percentage of kdrl :mCherry + ; runx1 :eGFP + hemogenic endothelial cells per embryo at 28 hpf (right) ( n = 4, mean ± SD, Student's t test). L) Confocal imaging of kdrl :mCherry + ; cmyb :eGFP + hemogenic endothelial cells in AGM at 48 hpf (left, white arrowheads). Graphs depicting the number of kdrl :mCherry + ; cmyb :eGFP + cells per embryo at 48 hpf (right) ( n = 5, mean ± SD, Student's t test). M) Flow cytometry plots of cd41 :eGFP low cells at 3 dpf (left). Graphs depicting the percentage of cd41 :eGFP low HSPCs per embryo at 3 dpf (right) ( n = 5, mean ± SD, Student's t test). N,O) WISH for runx1 N) and %phenotype distribution O) in embryos at 28 hpf as indicated ( n = 4, mean ± SD, one‐way ANOVA). ** p <0.01; *** p < 0.001.

    Article Snippet: KDM6A (Cell Signaling Technology, 33 510) and SOCS3 (Abcam, ab280884) antibody were applied to the sections and incubated overnight at 4 °C in a humidified chamber.

    Techniques: Flow Cytometry, Imaging

    Loss of Kdm6a deregulates inflammatory gene expression in ECs. A) Flowchart of sorting and RNA sequencing. B) Heatmap of differentially expressed genes in endothelial cells from kdm6a morphants and their WT controls at 28 hpf via RNA‐seq (cut‐off fold change 1.5, p < 0.05). C) Gene ontology (GO) enrichment analysis of biological processes based on differentially expressed genes identified through RNA‐seq. D) Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis based on differentially expressed genes identified through RNA‐seq. E) Gene set enrichment analysis (GESA) of RNA expression profiles in endothelial cells from kdm6a morphants and their WT controls at 28 hpf.

    Journal: Advanced Science

    Article Title: KDM6A Deficiency Induces Myeloid Bias and Promotes CMML‐Like Disease Through JAK/STAT3 Activation by Repressing SOCS3

    doi: 10.1002/advs.202413091

    Figure Lengend Snippet: Loss of Kdm6a deregulates inflammatory gene expression in ECs. A) Flowchart of sorting and RNA sequencing. B) Heatmap of differentially expressed genes in endothelial cells from kdm6a morphants and their WT controls at 28 hpf via RNA‐seq (cut‐off fold change 1.5, p < 0.05). C) Gene ontology (GO) enrichment analysis of biological processes based on differentially expressed genes identified through RNA‐seq. D) Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis based on differentially expressed genes identified through RNA‐seq. E) Gene set enrichment analysis (GESA) of RNA expression profiles in endothelial cells from kdm6a morphants and their WT controls at 28 hpf.

    Article Snippet: KDM6A (Cell Signaling Technology, 33 510) and SOCS3 (Abcam, ab280884) antibody were applied to the sections and incubated overnight at 4 °C in a humidified chamber.

    Techniques: Gene Expression, RNA Sequencing, RNA Expression

    Kdm6a regulates HSPC production via Syk‐associated inflammatory response in a H3K27me3‐dependent manner. A) Heatmap showing differential expression of inflammatory‐associated genes. B) Enrichment plot of the TNFα signaling via NF‐κB between differentially regulated genes in kdm6a morphants and their WT controls by GSEA. C,D) WISH for runx1 C) and %phenotype distribution D) in WT control, kdm6a mutants, and kdm6a mutants with ikbaa morphants at 28 hpf as indicated ( n = 4, mean ± SD, one‐way ANOVA). E) qPCR analysis of syk expression in sorted endothelial cells from WT control and kdm6a mutants at 28 hpf (n = 3, mean ± SD, Student's t test). F,G) WISH for runx1 F) and %phenotype distribution G) in WT control, kdm6a mutants, and kdm6a mutants with kdrl : syk constructs at 28 hpf ( n = 4, mean ± SD, one‐way ANOVA). H,I) WISH for runx1 H) and %phenotype distribution I) in WT control, kdm6a mutants, kdm6a mutants with kdm6a mRNA, and kdm6a mutants with kdm6a H1134A mRNA at 28 hpf ( n = 4, mean ± SD, one‐way ANOVA). J) ChIP‐qPCR analyses of syk promoter in kdm6a morphants and their WT controls by using an anti‐H3K27me3 antibody at 28 hpf (n = 3, mean ± SD, Student's t test). * p <0.05; ** p <0.01; *** p < 0.001.

    Journal: Advanced Science

    Article Title: KDM6A Deficiency Induces Myeloid Bias and Promotes CMML‐Like Disease Through JAK/STAT3 Activation by Repressing SOCS3

    doi: 10.1002/advs.202413091

    Figure Lengend Snippet: Kdm6a regulates HSPC production via Syk‐associated inflammatory response in a H3K27me3‐dependent manner. A) Heatmap showing differential expression of inflammatory‐associated genes. B) Enrichment plot of the TNFα signaling via NF‐κB between differentially regulated genes in kdm6a morphants and their WT controls by GSEA. C,D) WISH for runx1 C) and %phenotype distribution D) in WT control, kdm6a mutants, and kdm6a mutants with ikbaa morphants at 28 hpf as indicated ( n = 4, mean ± SD, one‐way ANOVA). E) qPCR analysis of syk expression in sorted endothelial cells from WT control and kdm6a mutants at 28 hpf (n = 3, mean ± SD, Student's t test). F,G) WISH for runx1 F) and %phenotype distribution G) in WT control, kdm6a mutants, and kdm6a mutants with kdrl : syk constructs at 28 hpf ( n = 4, mean ± SD, one‐way ANOVA). H,I) WISH for runx1 H) and %phenotype distribution I) in WT control, kdm6a mutants, kdm6a mutants with kdm6a mRNA, and kdm6a mutants with kdm6a H1134A mRNA at 28 hpf ( n = 4, mean ± SD, one‐way ANOVA). J) ChIP‐qPCR analyses of syk promoter in kdm6a morphants and their WT controls by using an anti‐H3K27me3 antibody at 28 hpf (n = 3, mean ± SD, Student's t test). * p <0.05; ** p <0.01; *** p < 0.001.

    Article Snippet: KDM6A (Cell Signaling Technology, 33 510) and SOCS3 (Abcam, ab280884) antibody were applied to the sections and incubated overnight at 4 °C in a humidified chamber.

    Techniques: Quantitative Proteomics, Control, Expressing, Construct, ChIP-qPCR

    Haploinsufficiency of Kdm6a has skewed myelopoiesis in embryonic and adult zebrafish. A,B) WISH for mfap4 A) and %phenotype distribution B) in embryos at 3 dpf as indicated (n = 4, mean ± SD, Student's t test). C,D) WISH for lyz C) and %phenotype distribution D) in embryos at 3 dpf as indicated ( n = 4, mean ± SD, Student's t test). E,F) WISH for hbae1 E) and %phenotype distribution F) in embryos at 3 dpf as indicated (n = 4, mean ± SD, Student's t test). G–L) qPCR analysis of mfap4 , mpeg1 , lyz , mpx , hbae1 , and alas2 expression in WT control and kdm6a +/− mutant embryos at 3 dpf (n = 3, mean ± SD, Student's t test). M) qPCR analysis of lineage differentiation‐related transcription factors expression including cebpd , cebpg , cebpz , irf8 , gata1a , and ccr9a in cd41 :eGFP low HSPCs isolated from WT control or kdm6a +/− mutant embryos (n = 3, mean ± SD, one‐way ANOVA). N‐N’) Hematoxylin and eosin staining of paraffin‐embedded sections of kidney from representative WT control or kdm6a +/− mutant adults. O‐O’) Hematoxylin and eosin staining of paraffin‐embedded sections of spleen from representative WT control or kdm6a +/− mutant adults. P‐P’) May–Grünwald–Giemsa staining of whole kidney marrow (KM) cells presentative 3 kdm6a +/− fish with CMML‐like phenotypes (totally 12 kdm6a +/− adults were used for experiment). * p <0.05; ** p <0.01; *** p < 0.001.

    Journal: Advanced Science

    Article Title: KDM6A Deficiency Induces Myeloid Bias and Promotes CMML‐Like Disease Through JAK/STAT3 Activation by Repressing SOCS3

    doi: 10.1002/advs.202413091

    Figure Lengend Snippet: Haploinsufficiency of Kdm6a has skewed myelopoiesis in embryonic and adult zebrafish. A,B) WISH for mfap4 A) and %phenotype distribution B) in embryos at 3 dpf as indicated (n = 4, mean ± SD, Student's t test). C,D) WISH for lyz C) and %phenotype distribution D) in embryos at 3 dpf as indicated ( n = 4, mean ± SD, Student's t test). E,F) WISH for hbae1 E) and %phenotype distribution F) in embryos at 3 dpf as indicated (n = 4, mean ± SD, Student's t test). G–L) qPCR analysis of mfap4 , mpeg1 , lyz , mpx , hbae1 , and alas2 expression in WT control and kdm6a +/− mutant embryos at 3 dpf (n = 3, mean ± SD, Student's t test). M) qPCR analysis of lineage differentiation‐related transcription factors expression including cebpd , cebpg , cebpz , irf8 , gata1a , and ccr9a in cd41 :eGFP low HSPCs isolated from WT control or kdm6a +/− mutant embryos (n = 3, mean ± SD, one‐way ANOVA). N‐N’) Hematoxylin and eosin staining of paraffin‐embedded sections of kidney from representative WT control or kdm6a +/− mutant adults. O‐O’) Hematoxylin and eosin staining of paraffin‐embedded sections of spleen from representative WT control or kdm6a +/− mutant adults. P‐P’) May–Grünwald–Giemsa staining of whole kidney marrow (KM) cells presentative 3 kdm6a +/− fish with CMML‐like phenotypes (totally 12 kdm6a +/− adults were used for experiment). * p <0.05; ** p <0.01; *** p < 0.001.

    Article Snippet: KDM6A (Cell Signaling Technology, 33 510) and SOCS3 (Abcam, ab280884) antibody were applied to the sections and incubated overnight at 4 °C in a humidified chamber.

    Techniques: Expressing, Control, Mutagenesis, Isolation, Staining

    Haploinsufficiency of Kdm6a dysregulates aging related gene expression in HSPCs. A) Flowchart of sorting and RNA sequencing. B) Heatmap of differentially expressed genes in HSPCs from kdm6a +/− mutants and their WT controls at 3 dpf via RNA‐seq (cut‐off fold change 1.5, p < 0.05). C) Gene ontology (GO) enrichment analysis of biological processes based on differentially expressed genes identified through RNA‐seq. D) Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis based on differentially expressed genes identified through RNA‐seq. E) Gene set enrichment analysis (GESA) of RNA expression profiles in HSPCs from kdm6a +/− mutants and their WT controls at 3 dpf.

    Journal: Advanced Science

    Article Title: KDM6A Deficiency Induces Myeloid Bias and Promotes CMML‐Like Disease Through JAK/STAT3 Activation by Repressing SOCS3

    doi: 10.1002/advs.202413091

    Figure Lengend Snippet: Haploinsufficiency of Kdm6a dysregulates aging related gene expression in HSPCs. A) Flowchart of sorting and RNA sequencing. B) Heatmap of differentially expressed genes in HSPCs from kdm6a +/− mutants and their WT controls at 3 dpf via RNA‐seq (cut‐off fold change 1.5, p < 0.05). C) Gene ontology (GO) enrichment analysis of biological processes based on differentially expressed genes identified through RNA‐seq. D) Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis based on differentially expressed genes identified through RNA‐seq. E) Gene set enrichment analysis (GESA) of RNA expression profiles in HSPCs from kdm6a +/− mutants and their WT controls at 3 dpf.

    Article Snippet: KDM6A (Cell Signaling Technology, 33 510) and SOCS3 (Abcam, ab280884) antibody were applied to the sections and incubated overnight at 4 °C in a humidified chamber.

    Techniques: Gene Expression, RNA Sequencing, RNA Expression

    Haploinsufficiency of Kdm6a promotes myeloid‐biased hematopoiesis through repressing socs3a and activating Jak/Stat3 signaling. A) Heatmaps showing the binding signals of ATAC at the promoter regions (TSS±2kb) in cd41 :eGFP low HSPCs at 3 dpf from WT control and kdm6a +/− mutants. B) Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis based on genes with differentially accessible chromatin at promoter regions. C) Venn plot showing the overlap of genes with specific accessible chromatin at promoter regions and downregulated expression in kdm6a +/− HSPCs. Genes for ATAC‐seq were assigned by differentially accessible chromatin at promoter regions. D) Genome browser tracks for ATAC enrichment signals across the socs3a locus in WT control and kdm6a +/− mutants. E) qPCR analysis of socs3a expression in sorted HSPCs from WT control and kdm6a +/− mutants at 3 dpf (n = 3, mean ± SD, Student's t test). F,G) WISH for runx1 F) and %phenotype distribution G) in WT control, kdm6a +/− mutants, and kdm6a +/− mutants with socs3a mRNA at 3 dpf ( n = 4, mean ± SD, one‐way ANOVA). (H‐I) WISH for mfap4 H) and %phenotype distribution I) in WT control, kdm6a +/− mutants, and kdm6a +/− mutants with socs3a mRNA at 3 dpf ( n = 4, mean ± SD, one‐way ANOVA). J) Enrichment plot of the STAT3 targets between differentially regulated genes in kdm6a +/− HSPCs and their WT controls by GSEA. K) qPCR analysis of STAT3 targets including ddx21 , c1qbp , and slc25a12 in cd41 :eGFP low HSPCs isolated from WT control and kdm6a +/− mutant embryos (n = 3, mean ± SD, one‐way ANOVA). (L‐M) WISH for lyz L) and %phenotype distribution M) in WT control, kdm6a +/− mutants, kdm6a +/− mutants with ruxolitinib, kdm6a +/− mutants with static, and kdm6a +/− mutants with baricitinib at 3 dpf ( n = 3, mean ± SD, one‐way ANOVA). N,O) WISH for mfap4 N) and %phenotype distribution O) in WT control, kdm6a +/− mutants, kdm6a +/− mutants with ruxolitinib, kdm6a +/− mutants with static, and kdm6a +/− mutants with baricitinib at 3 dpf (n = 3, mean ± SD, one‐way ANOVA). * p <0.05; ** p <0.01; *** p < 0.001.

    Journal: Advanced Science

    Article Title: KDM6A Deficiency Induces Myeloid Bias and Promotes CMML‐Like Disease Through JAK/STAT3 Activation by Repressing SOCS3

    doi: 10.1002/advs.202413091

    Figure Lengend Snippet: Haploinsufficiency of Kdm6a promotes myeloid‐biased hematopoiesis through repressing socs3a and activating Jak/Stat3 signaling. A) Heatmaps showing the binding signals of ATAC at the promoter regions (TSS±2kb) in cd41 :eGFP low HSPCs at 3 dpf from WT control and kdm6a +/− mutants. B) Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis based on genes with differentially accessible chromatin at promoter regions. C) Venn plot showing the overlap of genes with specific accessible chromatin at promoter regions and downregulated expression in kdm6a +/− HSPCs. Genes for ATAC‐seq were assigned by differentially accessible chromatin at promoter regions. D) Genome browser tracks for ATAC enrichment signals across the socs3a locus in WT control and kdm6a +/− mutants. E) qPCR analysis of socs3a expression in sorted HSPCs from WT control and kdm6a +/− mutants at 3 dpf (n = 3, mean ± SD, Student's t test). F,G) WISH for runx1 F) and %phenotype distribution G) in WT control, kdm6a +/− mutants, and kdm6a +/− mutants with socs3a mRNA at 3 dpf ( n = 4, mean ± SD, one‐way ANOVA). (H‐I) WISH for mfap4 H) and %phenotype distribution I) in WT control, kdm6a +/− mutants, and kdm6a +/− mutants with socs3a mRNA at 3 dpf ( n = 4, mean ± SD, one‐way ANOVA). J) Enrichment plot of the STAT3 targets between differentially regulated genes in kdm6a +/− HSPCs and their WT controls by GSEA. K) qPCR analysis of STAT3 targets including ddx21 , c1qbp , and slc25a12 in cd41 :eGFP low HSPCs isolated from WT control and kdm6a +/− mutant embryos (n = 3, mean ± SD, one‐way ANOVA). (L‐M) WISH for lyz L) and %phenotype distribution M) in WT control, kdm6a +/− mutants, kdm6a +/− mutants with ruxolitinib, kdm6a +/− mutants with static, and kdm6a +/− mutants with baricitinib at 3 dpf ( n = 3, mean ± SD, one‐way ANOVA). N,O) WISH for mfap4 N) and %phenotype distribution O) in WT control, kdm6a +/− mutants, kdm6a +/− mutants with ruxolitinib, kdm6a +/− mutants with static, and kdm6a +/− mutants with baricitinib at 3 dpf (n = 3, mean ± SD, one‐way ANOVA). * p <0.05; ** p <0.01; *** p < 0.001.

    Article Snippet: KDM6A (Cell Signaling Technology, 33 510) and SOCS3 (Abcam, ab280884) antibody were applied to the sections and incubated overnight at 4 °C in a humidified chamber.

    Techniques: Binding Assay, Control, Expressing, Isolation, Mutagenesis

    KDM6A/SOCS3/p‐STAT3 pathway is conserved in human HSPCs. A) Immunohistochemical staining of KDM6A on BM biopsies. B) Immunohistochemical staining of SOCS3 on BM biopsies. C) The percentage of KDM6A + and SOCS3 + cells in immunohistochemical staining. D) The H‐score of KDM6A + and SOCS3 + cells immunohistochemical staining. E) Correlation analysis of the percentage of KDM6A + (C, left) and SOCS3 + (C, right) cells in CMML specimens. F) Correlation analysis of the H‐score of KDM6A + (D, left) and SOCS3 + (D, right) cells in CMML specimens. G) Schematic representation of human HSPC enrichment, lentiviral infection, and in vitro stimulation experiments conducted in this study. H) qPCR analysis of genes expression including SOCS3 , DDX21 , C1QBP , SLC25A12 , CEBPG , CEBPZ , and IRF8 in human HSPCs after GM‐CSF stimulation ( n = 3, mean ± SD, Student's t test). I) Western Blot analysis of KDM6A, and H3K27me3 in human HSPCs after GM‐CSF stimulation. (J) Western Blot analysis of γH2AX, STAT3, and p‐STAT3 Y705 in human HSPCs after GM‐CSF stimulation. * p <0.05; ** p <0.01; *** p < 0.001.

    Journal: Advanced Science

    Article Title: KDM6A Deficiency Induces Myeloid Bias and Promotes CMML‐Like Disease Through JAK/STAT3 Activation by Repressing SOCS3

    doi: 10.1002/advs.202413091

    Figure Lengend Snippet: KDM6A/SOCS3/p‐STAT3 pathway is conserved in human HSPCs. A) Immunohistochemical staining of KDM6A on BM biopsies. B) Immunohistochemical staining of SOCS3 on BM biopsies. C) The percentage of KDM6A + and SOCS3 + cells in immunohistochemical staining. D) The H‐score of KDM6A + and SOCS3 + cells immunohistochemical staining. E) Correlation analysis of the percentage of KDM6A + (C, left) and SOCS3 + (C, right) cells in CMML specimens. F) Correlation analysis of the H‐score of KDM6A + (D, left) and SOCS3 + (D, right) cells in CMML specimens. G) Schematic representation of human HSPC enrichment, lentiviral infection, and in vitro stimulation experiments conducted in this study. H) qPCR analysis of genes expression including SOCS3 , DDX21 , C1QBP , SLC25A12 , CEBPG , CEBPZ , and IRF8 in human HSPCs after GM‐CSF stimulation ( n = 3, mean ± SD, Student's t test). I) Western Blot analysis of KDM6A, and H3K27me3 in human HSPCs after GM‐CSF stimulation. (J) Western Blot analysis of γH2AX, STAT3, and p‐STAT3 Y705 in human HSPCs after GM‐CSF stimulation. * p <0.05; ** p <0.01; *** p < 0.001.

    Article Snippet: KDM6A (Cell Signaling Technology, 33 510) and SOCS3 (Abcam, ab280884) antibody were applied to the sections and incubated overnight at 4 °C in a humidified chamber.

    Techniques: Immunohistochemical staining, Staining, Infection, In Vitro, Expressing, Western Blot

    Primary antibodies used for indirect immunofluorescence

    Journal: Epigenetics & Chromatin

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes

    doi: 10.1186/s13072-025-00583-9

    Figure Lengend Snippet: Primary antibodies used for indirect immunofluorescence

    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Techniques: Biomarker Discovery, Knock-Out, Knockdown, Mutagenesis, Membrane

    H3K27me3 peaks in primary follicle oocytes before declining coincident with increasing KDM6A in primary-secondary stage. ( A , C ) Representative IF images showing H3K27me3 (A) and KDM6A (C) (red, grey single channel) in primordial, primary, secondary, early antral and antral follicle oocytes from wildtype adult mouse ovaries. Images are representative of multiple planes in both ovaries from two to three biological replicates. The oocyte nucleus is defined by Lamin B1 (green, dashed lines), and DNA is shown by DAPI (blue). White boxes indicate regions shown at higher power in images on the right. Yellow arrowheads denote primordial follicle oocyte nuclei. Scale bars represent 50 μm (left), and 10 μm (middle, right). ( B , D ) Quantification of H3K27me3 (B) and KDM6A (D) within oocyte nuclei of primordial, primary, secondary, early antral and antral follicles from wildtype adult mouse ovaries. Values represent average nuclear staining intensity with average cytoplasmic fluorescence removed to correct for non-specific background staining. Error bars represent mean ± SD. (B) ns: not significant, * P < 0.05, *** P < 0.001, **** P < 0.0001, Kruskal-Wallis test with Dunn’s multiple comparisons test. N = 84 primordial follicle oocytes, N = 37 primary follicle oocytes, N = 32 secondary follicle oocytes, N = 11 early antral follicle oocytes, and N = 4 antral follicle oocytes, in total from three biological replicates. ( D ) Quantification of KDM6A within oocyte nuclei of primordial, primary, secondary, early antral and antral follicles from wildtype adult mouse ovaries. ns: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, one-way ANOVA plus Tukey’s multiple comparisons test. N = 17 primordial follicle oocytes, N = 23 primary follicle oocytes, N = 24 secondary follicle oocytes, N = 12 early antral follicle oocytes, and N = 5 antral follicle oocytes, in total from two biological replicates

    Journal: Epigenetics & Chromatin

    Article Title: Coordinated regulation of chromatin modifiers reflects organised epigenetic programming in mouse oocytes

    doi: 10.1186/s13072-025-00583-9

    Figure Lengend Snippet: H3K27me3 peaks in primary follicle oocytes before declining coincident with increasing KDM6A in primary-secondary stage. ( A , C ) Representative IF images showing H3K27me3 (A) and KDM6A (C) (red, grey single channel) in primordial, primary, secondary, early antral and antral follicle oocytes from wildtype adult mouse ovaries. Images are representative of multiple planes in both ovaries from two to three biological replicates. The oocyte nucleus is defined by Lamin B1 (green, dashed lines), and DNA is shown by DAPI (blue). White boxes indicate regions shown at higher power in images on the right. Yellow arrowheads denote primordial follicle oocyte nuclei. Scale bars represent 50 μm (left), and 10 μm (middle, right). ( B , D ) Quantification of H3K27me3 (B) and KDM6A (D) within oocyte nuclei of primordial, primary, secondary, early antral and antral follicles from wildtype adult mouse ovaries. Values represent average nuclear staining intensity with average cytoplasmic fluorescence removed to correct for non-specific background staining. Error bars represent mean ± SD. (B) ns: not significant, * P < 0.05, *** P < 0.001, **** P < 0.0001, Kruskal-Wallis test with Dunn’s multiple comparisons test. N = 84 primordial follicle oocytes, N = 37 primary follicle oocytes, N = 32 secondary follicle oocytes, N = 11 early antral follicle oocytes, and N = 4 antral follicle oocytes, in total from three biological replicates. ( D ) Quantification of KDM6A within oocyte nuclei of primordial, primary, secondary, early antral and antral follicles from wildtype adult mouse ovaries. ns: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, one-way ANOVA plus Tukey’s multiple comparisons test. N = 17 primordial follicle oocytes, N = 23 primary follicle oocytes, N = 24 secondary follicle oocytes, N = 12 early antral follicle oocytes, and N = 5 antral follicle oocytes, in total from two biological replicates

    Article Snippet: KDM6A , Rabbit , 1:400 , Validated in THP-1 cells with loss of KDM6A , Cell Signaling Technology, 33510S.

    Techniques: Staining, Fluorescence

    Kdm6a KO results in Xist downregulation. A Principal component analysis (PCA) based on expression of autosomal or X-linked genes in two Tsix -stop wt clones and two Kdm6a KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) before (D0) and after differentiation (D15). Only genes with a minimum of 2.5 CPM in 4 libraries were considered. PCA shows separation of clones based on Kdm6a KO status mainly after differentiation. PCA figures were generated by iDEP.95. B Plots of diploid gene expression from a representative autosome (Chr1, left) with similar gene content as the X, and from the X chromosome (right) in Tsix -stop wt and KO clones (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D15. Kdm6a and the XIC are highlighted. Log 2 TPM fold change (KO/wt) is shown. Genes with a decrease of expression in KO versus wt are in blue, and with an increase in red. C qRT-PCR of average Xist expression normalized to Actinβ during ES differentiation in three Tsix -stop wt samples, two Kdm6a ΔEΔE KO clones, and one Kdm6a ΔwtΔE clone. p-value is derived from comparing wt to Kdm6a ΔEΔE biological replicates. Divergence of Xist expression between wt and KO cells begins at D2 and is largest at D7 (p = 0.04). The Kdm6a wt/ΔE4 heterozygous KO clone behaves similarly to the homozygous clones. D qRT-PCR of Xist expression in E8 wt and KO clone (E8- Kdm6a ΔPΔP13 ). Expression is normalized to Actinβ . Xist is significantly lower at D2 and D4 in KO cells. P-values derived from two technical replicates of wt and KO cells (**p < 0.01;*p < 0.05). E RNA-FISH with a probe specific for Xist RNA labeled in green in Tsix -stop wt and KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ). Examples of nuclei are shown together with histograms for quantification of signal type (pinpoint, no signal or cloud). Xist clouds are present in 74% of D15 wt cells, but significantly decrease in KO clones (cloud compared to no signal; ***p < 0.00001 for T six-Kdm6a ΔEΔE17 and < 0.0001 for Tsix-Kdm6a ΔEΔE21 fisher’s exact test), while the frequency of pinpoint Xist signals increases in KO clones (pinpoint in wt compared to pinpoint in KO; ***p < 0.00001 for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 fisher’s exact test). Nuclei are counterstained with Hoechst 33,342. Scale bar = 10 µm

    Journal: Biology of Sex Differences

    Article Title: KDM6A facilitates Xist upregulation at the onset of X inactivation

    doi: 10.1186/s13293-024-00683-3

    Figure Lengend Snippet: Kdm6a KO results in Xist downregulation. A Principal component analysis (PCA) based on expression of autosomal or X-linked genes in two Tsix -stop wt clones and two Kdm6a KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) before (D0) and after differentiation (D15). Only genes with a minimum of 2.5 CPM in 4 libraries were considered. PCA shows separation of clones based on Kdm6a KO status mainly after differentiation. PCA figures were generated by iDEP.95. B Plots of diploid gene expression from a representative autosome (Chr1, left) with similar gene content as the X, and from the X chromosome (right) in Tsix -stop wt and KO clones (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D15. Kdm6a and the XIC are highlighted. Log 2 TPM fold change (KO/wt) is shown. Genes with a decrease of expression in KO versus wt are in blue, and with an increase in red. C qRT-PCR of average Xist expression normalized to Actinβ during ES differentiation in three Tsix -stop wt samples, two Kdm6a ΔEΔE KO clones, and one Kdm6a ΔwtΔE clone. p-value is derived from comparing wt to Kdm6a ΔEΔE biological replicates. Divergence of Xist expression between wt and KO cells begins at D2 and is largest at D7 (p = 0.04). The Kdm6a wt/ΔE4 heterozygous KO clone behaves similarly to the homozygous clones. D qRT-PCR of Xist expression in E8 wt and KO clone (E8- Kdm6a ΔPΔP13 ). Expression is normalized to Actinβ . Xist is significantly lower at D2 and D4 in KO cells. P-values derived from two technical replicates of wt and KO cells (**p < 0.01;*p < 0.05). E RNA-FISH with a probe specific for Xist RNA labeled in green in Tsix -stop wt and KO clones ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ). Examples of nuclei are shown together with histograms for quantification of signal type (pinpoint, no signal or cloud). Xist clouds are present in 74% of D15 wt cells, but significantly decrease in KO clones (cloud compared to no signal; ***p < 0.00001 for T six-Kdm6a ΔEΔE17 and < 0.0001 for Tsix-Kdm6a ΔEΔE21 fisher’s exact test), while the frequency of pinpoint Xist signals increases in KO clones (pinpoint in wt compared to pinpoint in KO; ***p < 0.00001 for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 fisher’s exact test). Nuclei are counterstained with Hoechst 33,342. Scale bar = 10 µm

    Article Snippet: Cut&Run for KDM6A was done in wt cells using an antibody against KDM6A (Cell signaling #33510) according to the manufacturer’s protocol (EpiCypher) except with mild fixation (0.01% formaldehyde for 1 min).

    Techniques: Expressing, Clone Assay, Generated, Gene Expression, Quantitative RT-PCR, Derivative Assay, Labeling

    Kdm6a KO leads to increased gene expression from the Xi in differentiated cells. A Allelic X-linked gene expression ratios (129:cast) in Tsix -stop wt, CRISPR control, and KO cells ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D0 and D15 of differentiation. Ratios between expressed X-linked genes (> 1TPM) are shown. There is increased gene expression from the 129 X-chromosome (Xi) following Kdm6a KO differentiation. B, C Plots of expression from the Xi (B) and the Xa (C) (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D15. Kdm6a and the XIC are highlighted. Log 2 TPM fold change (KO/wt) is shown. Genes with a decrease of expression in KO versus wt are in blue, and with an increase in red. Gene expression from the Xi is selectively increased following KO. D Top, scatter plots of average log 2 expression between Tsix- stop wt and KO clones (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D15 for X-linked genes categorized as those silenced early, silenced at an intermediate time, silenced late, and not silenced (escape) during XCI [ , ]. KO clones show higher expression of genes in each category compared to wt. n indicates the number of genes in each category. Bottom, histograms of median allelic expression of genes silenced at different times during XCI and of genes that escape XCI is increased in Tsix- stop KO clones compared to wt and CRISPR controls for all categories, while there is little change in expression from the Xa. n indicates the number of genes in each category. E Expression fold changes of individual escape genes from the Xi in Tsix- stop KO clones (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) compared to wt at D15. Most genes show increased expression

    Journal: Biology of Sex Differences

    Article Title: KDM6A facilitates Xist upregulation at the onset of X inactivation

    doi: 10.1186/s13293-024-00683-3

    Figure Lengend Snippet: Kdm6a KO leads to increased gene expression from the Xi in differentiated cells. A Allelic X-linked gene expression ratios (129:cast) in Tsix -stop wt, CRISPR control, and KO cells ( Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D0 and D15 of differentiation. Ratios between expressed X-linked genes (> 1TPM) are shown. There is increased gene expression from the 129 X-chromosome (Xi) following Kdm6a KO differentiation. B, C Plots of expression from the Xi (B) and the Xa (C) (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D15. Kdm6a and the XIC are highlighted. Log 2 TPM fold change (KO/wt) is shown. Genes with a decrease of expression in KO versus wt are in blue, and with an increase in red. Gene expression from the Xi is selectively increased following KO. D Top, scatter plots of average log 2 expression between Tsix- stop wt and KO clones (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) at D15 for X-linked genes categorized as those silenced early, silenced at an intermediate time, silenced late, and not silenced (escape) during XCI [ , ]. KO clones show higher expression of genes in each category compared to wt. n indicates the number of genes in each category. Bottom, histograms of median allelic expression of genes silenced at different times during XCI and of genes that escape XCI is increased in Tsix- stop KO clones compared to wt and CRISPR controls for all categories, while there is little change in expression from the Xa. n indicates the number of genes in each category. E Expression fold changes of individual escape genes from the Xi in Tsix- stop KO clones (average for Tsix-Kdm6a ΔEΔE17 and Tsix-Kdm6a ΔEΔE21 ) compared to wt at D15. Most genes show increased expression

    Article Snippet: Cut&Run for KDM6A was done in wt cells using an antibody against KDM6A (Cell signaling #33510) according to the manufacturer’s protocol (EpiCypher) except with mild fixation (0.01% formaldehyde for 1 min).

    Techniques: Gene Expression, CRISPR, Control, Expressing, Clone Assay

    Kdm6a KO leads to increased H3K27me3 at Xist specifically on the Xi. A IGV browser views of gene expression (exp) and H3K27me3 peaks across the 129 and cast X chromosomes obtained by allelic analysis in Tsix- stop wt and KO cells ( Tsix-Kdm6a ΔEΔE17 ) at D0 and D15. The 129 profile is in pink (wt) or purple (KO) and the cast in light blue (wt) or dark blue (KO). XCI is skewed in Tsix -stop cells so that the 129 X will become the Xi and the cast X the Xa. The number of peaks for each condition and time point are in parentheses. There is a marked increase in Xist expression from the Xi in wt cells but not in KO cells at D15. H3K27me3 levels increase following differentiation on the Xi in wt and Kdm6a KO cells. B Same analysis as in (A) , but zoomed in on the Xist gene. Allelic RNA-seq tracks of gene expression show very low expression from the 129 Xi allele in KO cells. A correlated Xi-specific increase in H3K27me3 enrichment at the Xist promoter is seen as an increase in peaks. No expression of Xist is observed from the cast allele, which is the Xa. The scales of the profiles are indicated in the upper right corners

    Journal: Biology of Sex Differences

    Article Title: KDM6A facilitates Xist upregulation at the onset of X inactivation

    doi: 10.1186/s13293-024-00683-3

    Figure Lengend Snippet: Kdm6a KO leads to increased H3K27me3 at Xist specifically on the Xi. A IGV browser views of gene expression (exp) and H3K27me3 peaks across the 129 and cast X chromosomes obtained by allelic analysis in Tsix- stop wt and KO cells ( Tsix-Kdm6a ΔEΔE17 ) at D0 and D15. The 129 profile is in pink (wt) or purple (KO) and the cast in light blue (wt) or dark blue (KO). XCI is skewed in Tsix -stop cells so that the 129 X will become the Xi and the cast X the Xa. The number of peaks for each condition and time point are in parentheses. There is a marked increase in Xist expression from the Xi in wt cells but not in KO cells at D15. H3K27me3 levels increase following differentiation on the Xi in wt and Kdm6a KO cells. B Same analysis as in (A) , but zoomed in on the Xist gene. Allelic RNA-seq tracks of gene expression show very low expression from the 129 Xi allele in KO cells. A correlated Xi-specific increase in H3K27me3 enrichment at the Xist promoter is seen as an increase in peaks. No expression of Xist is observed from the cast allele, which is the Xa. The scales of the profiles are indicated in the upper right corners

    Article Snippet: Cut&Run for KDM6A was done in wt cells using an antibody against KDM6A (Cell signaling #33510) according to the manufacturer’s protocol (EpiCypher) except with mild fixation (0.01% formaldehyde for 1 min).

    Techniques: Gene Expression, Expressing, RNA Sequencing