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immunoprecipitation reactions  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc immunoprecipitation reactions
    a , b Representative images and analysis of INCENP and CDCA8 protein expression levels after YTHDF3 knockdown in PDX-derived organoids (PDXO) by immunofluorescence. Scale bar = 10 μm. Mean protein fluorescence intensity was determined using ImageJ software. c , d Gene plots illustrating YTHDF3 binding to INCENP and CDCA8 mRNAs, as measured by RIP-seq. The normalized read distribution: input (yellow) and YTHDF3 (purple) along the mRNAs. e RNA <t>immunoprecipitation-qPCR</t> (RIP-qPCR) highlighting the indicated mRNA transcripts bound by YTHDF3 in ESCC cells. f , g Representative immunofluorescence staining of newly synthesized proteins and statistical analysis in KYSE30 and KYSE150 cells after YTHDF3 overexpression. Relative fluorescence intensity was determined using ImageJ software. Puromycin staining (green) indicates newly synthesized proteins; nuclei are stained with DAPI (blue). Scale bar = 50 μm. h , i Mock or YTHDF3 knockdown KYSE30 and KYSE150 cells were transfected with pmirGLO-INCENP or pmirGLO-CDCA8 reporters for 48 h. Translation efficiency of INCENP or CDCA8 is defined as the quotient of reporter protein production (F-luc/R-luc). j , k Co-immunoprecipitation of endogenous YTHDF3 and eIF3A in ESCC cells. Representative immunoblots shown in figures were repeated three times independently with similar results. l , m In situ detection and quantification of YTHDF3–eIF3A interactions in the indicated ESCC cell lines were measured using PLA assays, and puncta per cell were determined using ImageJ software. Scale bar = 10 μm, n = 5 biological replicates. n RIP-qPCR illustrating the association of eIF3A with the indicated transcripts in YTHDF3 knockdown KYSE30 cells. o RIP-qPCR illustrating the association of eIF3A with the indicated transcripts in Ebselen-treated KYSE30 cells. RIP-qPCR illustrating the association of YTHDF3 ( p ) and eIF3A ( q ) with the indicated transcripts in METTL3 knockdown KYSE30 cells. Data were analyzed by two-tailed unpaired t-test ( b , e , g , h , i , m ) or one-way ANOVA ( n , o , p , q ). n = 3 biological replicates ( b , e , g , h , i , n , o , p , q ). Data were presented as means ± SD. Source data are provided as a file.
    Immunoprecipitation Reactions, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "INCENP and CDCA8 predict neoadjuvant chemotherapy response and outcomes in esophageal squamous cell carcinoma"

    Article Title: INCENP and CDCA8 predict neoadjuvant chemotherapy response and outcomes in esophageal squamous cell carcinoma

    Journal: Nature Communications

    doi: 10.1038/s41467-026-68371-x

    a , b Representative images and analysis of INCENP and CDCA8 protein expression levels after YTHDF3 knockdown in PDX-derived organoids (PDXO) by immunofluorescence. Scale bar = 10 μm. Mean protein fluorescence intensity was determined using ImageJ software. c , d Gene plots illustrating YTHDF3 binding to INCENP and CDCA8 mRNAs, as measured by RIP-seq. The normalized read distribution: input (yellow) and YTHDF3 (purple) along the mRNAs. e RNA immunoprecipitation-qPCR (RIP-qPCR) highlighting the indicated mRNA transcripts bound by YTHDF3 in ESCC cells. f , g Representative immunofluorescence staining of newly synthesized proteins and statistical analysis in KYSE30 and KYSE150 cells after YTHDF3 overexpression. Relative fluorescence intensity was determined using ImageJ software. Puromycin staining (green) indicates newly synthesized proteins; nuclei are stained with DAPI (blue). Scale bar = 50 μm. h , i Mock or YTHDF3 knockdown KYSE30 and KYSE150 cells were transfected with pmirGLO-INCENP or pmirGLO-CDCA8 reporters for 48 h. Translation efficiency of INCENP or CDCA8 is defined as the quotient of reporter protein production (F-luc/R-luc). j , k Co-immunoprecipitation of endogenous YTHDF3 and eIF3A in ESCC cells. Representative immunoblots shown in figures were repeated three times independently with similar results. l , m In situ detection and quantification of YTHDF3–eIF3A interactions in the indicated ESCC cell lines were measured using PLA assays, and puncta per cell were determined using ImageJ software. Scale bar = 10 μm, n = 5 biological replicates. n RIP-qPCR illustrating the association of eIF3A with the indicated transcripts in YTHDF3 knockdown KYSE30 cells. o RIP-qPCR illustrating the association of eIF3A with the indicated transcripts in Ebselen-treated KYSE30 cells. RIP-qPCR illustrating the association of YTHDF3 ( p ) and eIF3A ( q ) with the indicated transcripts in METTL3 knockdown KYSE30 cells. Data were analyzed by two-tailed unpaired t-test ( b , e , g , h , i , m ) or one-way ANOVA ( n , o , p , q ). n = 3 biological replicates ( b , e , g , h , i , n , o , p , q ). Data were presented as means ± SD. Source data are provided as a file.
    Figure Legend Snippet: a , b Representative images and analysis of INCENP and CDCA8 protein expression levels after YTHDF3 knockdown in PDX-derived organoids (PDXO) by immunofluorescence. Scale bar = 10 μm. Mean protein fluorescence intensity was determined using ImageJ software. c , d Gene plots illustrating YTHDF3 binding to INCENP and CDCA8 mRNAs, as measured by RIP-seq. The normalized read distribution: input (yellow) and YTHDF3 (purple) along the mRNAs. e RNA immunoprecipitation-qPCR (RIP-qPCR) highlighting the indicated mRNA transcripts bound by YTHDF3 in ESCC cells. f , g Representative immunofluorescence staining of newly synthesized proteins and statistical analysis in KYSE30 and KYSE150 cells after YTHDF3 overexpression. Relative fluorescence intensity was determined using ImageJ software. Puromycin staining (green) indicates newly synthesized proteins; nuclei are stained with DAPI (blue). Scale bar = 50 μm. h , i Mock or YTHDF3 knockdown KYSE30 and KYSE150 cells were transfected with pmirGLO-INCENP or pmirGLO-CDCA8 reporters for 48 h. Translation efficiency of INCENP or CDCA8 is defined as the quotient of reporter protein production (F-luc/R-luc). j , k Co-immunoprecipitation of endogenous YTHDF3 and eIF3A in ESCC cells. Representative immunoblots shown in figures were repeated three times independently with similar results. l , m In situ detection and quantification of YTHDF3–eIF3A interactions in the indicated ESCC cell lines were measured using PLA assays, and puncta per cell were determined using ImageJ software. Scale bar = 10 μm, n = 5 biological replicates. n RIP-qPCR illustrating the association of eIF3A with the indicated transcripts in YTHDF3 knockdown KYSE30 cells. o RIP-qPCR illustrating the association of eIF3A with the indicated transcripts in Ebselen-treated KYSE30 cells. RIP-qPCR illustrating the association of YTHDF3 ( p ) and eIF3A ( q ) with the indicated transcripts in METTL3 knockdown KYSE30 cells. Data were analyzed by two-tailed unpaired t-test ( b , e , g , h , i , m ) or one-way ANOVA ( n , o , p , q ). n = 3 biological replicates ( b , e , g , h , i , n , o , p , q ). Data were presented as means ± SD. Source data are provided as a file.

    Techniques Used: Expressing, Knockdown, Derivative Assay, Immunofluorescence, Fluorescence, Software, Binding Assay, RNA Immunoprecipitation, Staining, Synthesized, Over Expression, Transfection, Immunoprecipitation, Western Blot, In Situ, Two Tailed Test

    Related Articles

    other:

    Article Title: Neuronal hyperactivity becomes mTORC1 independent due to transcriptional changes in tuberous sclerosis complex disease models
    Article Snippet: hiPSC-derived neurons at the indicated day of differentiation were washed with PBS and collected in RIPA buffer with protease inhibitors (Thermo Fisher Scientific, A32955).

    Incubation:

    Article Title: Regulation and Function of the HPV16 CircE7 RNA
    Article Snippet: For western blot analysis, cells were lysed in Cell Lysis Buffer (Cell Signaling Technologies, 5872S), separated on SDS-PAGE gels, and transferred to PVDF membranes. .. Membranes were incubated overnight at 4°C with the following primary antibodies: YTHDC1 (Cell Signaling, 77422), YTHDF3 (Cell Signaling, 24206), YTHDC2 (Cell Signaling, 46324), FLAG (Sigma, A8592), GAPDH (Santa Cruz, sc-32233), HSP90 (Cell Signaling, 4877S). .. Western blots were then incubated with the appropriate HRP-conjugated secondary antibodies (Cell Signaling, 7076S and 7074S) and ECL substrate (Perkin Elmer, 50-904-9323) for signal detection.

    Article Title: INCENP and CDCA8 predict neoadjuvant chemotherapy response and outcomes in esophageal squamous cell carcinoma
    Article Snippet: .. Ten percent (10%) of the lysate was stored as “input”, 80% was used in immunoprecipitation reactions with anti-YTHDF3 antibody (Abcam, ab220161), and the remaining 10% was incubated with rabbit IgG (Cell Signaling Technology) as a negative control and named “IgG”, respectively. .. The RNA of input and IP was extracted using TRIzol reagent (Invitrogen, cat. NO 15596026).

    Article Title: INCENP and CDCA8 predict neoadjuvant chemotherapy response and outcomes in esophageal squamous cell carcinoma.
    Article Snippet: .. Ten percent (10%) of the lysate was stored as “input”, 80% was used in immunoprecipitation reactions with anti-YTHDF3 antibody (Abcam, ab220161), and the remaining 10% was incubated with rabbit IgG (Cell Signaling Technology) as a negative control and named “IgG”, respectively. .. The RNA of input and IP was extracted using TRIzol reagent (Invitrogen, cat. NO 15596026).

    Immunoprecipitation:

    Article Title: INCENP and CDCA8 predict neoadjuvant chemotherapy response and outcomes in esophageal squamous cell carcinoma
    Article Snippet: .. Ten percent (10%) of the lysate was stored as “input”, 80% was used in immunoprecipitation reactions with anti-YTHDF3 antibody (Abcam, ab220161), and the remaining 10% was incubated with rabbit IgG (Cell Signaling Technology) as a negative control and named “IgG”, respectively. .. The RNA of input and IP was extracted using TRIzol reagent (Invitrogen, cat. NO 15596026).

    Article Title: INCENP and CDCA8 predict neoadjuvant chemotherapy response and outcomes in esophageal squamous cell carcinoma.
    Article Snippet: .. Ten percent (10%) of the lysate was stored as “input”, 80% was used in immunoprecipitation reactions with anti-YTHDF3 antibody (Abcam, ab220161), and the remaining 10% was incubated with rabbit IgG (Cell Signaling Technology) as a negative control and named “IgG”, respectively. .. The RNA of input and IP was extracted using TRIzol reagent (Invitrogen, cat. NO 15596026).

    Negative Control:

    Article Title: INCENP and CDCA8 predict neoadjuvant chemotherapy response and outcomes in esophageal squamous cell carcinoma
    Article Snippet: .. Ten percent (10%) of the lysate was stored as “input”, 80% was used in immunoprecipitation reactions with anti-YTHDF3 antibody (Abcam, ab220161), and the remaining 10% was incubated with rabbit IgG (Cell Signaling Technology) as a negative control and named “IgG”, respectively. .. The RNA of input and IP was extracted using TRIzol reagent (Invitrogen, cat. NO 15596026).

    Article Title: INCENP and CDCA8 predict neoadjuvant chemotherapy response and outcomes in esophageal squamous cell carcinoma.
    Article Snippet: .. Ten percent (10%) of the lysate was stored as “input”, 80% was used in immunoprecipitation reactions with anti-YTHDF3 antibody (Abcam, ab220161), and the remaining 10% was incubated with rabbit IgG (Cell Signaling Technology) as a negative control and named “IgG”, respectively. .. The RNA of input and IP was extracted using TRIzol reagent (Invitrogen, cat. NO 15596026).



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    YTHDF3 is overexpressed in breast cancer tissues and cells. (A) YTHDF3 expression in breast cancer tissues and adjacent normal tissues in TCGA database; (B) YTHDF3 expression in different types of breast cancer in TCGA database; (C) Kaplan–Meier curves of OS in the TCGA dataset of all breast cancer patients with high and low YTHDF3 expression; (D) YTHDF3 expression in paired breast cancer tissues and adjacent normal tissues; (E) YTHDF3 expression of breast cancer and para‐cancerous tissues was evaluated by immunohistochemistry, ×100; (F) YTHDF3 protein expression in normal human breast cell line MCF‐10A and breast cancer cell lines MCF‐7, MDA‐MB‐231, SK‐BR‐3, ZR‐75‐1 and MDA‐MB‐453; (G) Relative YTHDF3 mRNA expression in normal human breast cell line MCF‐10A and breast cancer cell lines MCF‐7, MDA‐MB‐231, SK‐BR‐3, ZR‐75‐1 and MDA‐MB‐453.

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

    doi: 10.1111/jcmm.71105

    Figure Lengend Snippet: YTHDF3 is overexpressed in breast cancer tissues and cells. (A) YTHDF3 expression in breast cancer tissues and adjacent normal tissues in TCGA database; (B) YTHDF3 expression in different types of breast cancer in TCGA database; (C) Kaplan–Meier curves of OS in the TCGA dataset of all breast cancer patients with high and low YTHDF3 expression; (D) YTHDF3 expression in paired breast cancer tissues and adjacent normal tissues; (E) YTHDF3 expression of breast cancer and para‐cancerous tissues was evaluated by immunohistochemistry, ×100; (F) YTHDF3 protein expression in normal human breast cell line MCF‐10A and breast cancer cell lines MCF‐7, MDA‐MB‐231, SK‐BR‐3, ZR‐75‐1 and MDA‐MB‐453; (G) Relative YTHDF3 mRNA expression in normal human breast cell line MCF‐10A and breast cancer cell lines MCF‐7, MDA‐MB‐231, SK‐BR‐3, ZR‐75‐1 and MDA‐MB‐453.

    Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

    Techniques: Expressing, Immunohistochemistry

    Silence of YTHDF3 restrain cell proliferation and migration of breast cancer cells and arrested the cell cycle. (A) Relative expression of YTHDF3 mRNA and protein after transfection of shNC and shYTHDF3 in MDA‐MB‐231 and MCF‐7 cells; (B) the growth ability of MDA‐MB‐231 and MCF‐7 cells transfected with shNC and shYTHDF3 was detected by CCK‐8 assay; (C) colony formation assay was used to detect the proliferation ability of MDA‐MB‐231 and MCF‐7 cells in shNC and shYTHDF3 groups; (D) the cell cycle distribution of MDA‐MB‐231 and MCF‐7 cells in shNC and shYTHDF3 groups was detected by flow cytometry; (E) expression of cyclin D1 and CDK4 in shNC and shYTHDF3 in MDA‐MB‐231 and MCF‐7 cells; (F, G) transwell invasion assay was used to detect the migration ability of breast cancer cells in the shNC group and the shYTHDF3 group in MCF‐7 and MDA‐MB‐231 cells; (H) the expression of vimentin, MMP9, E‐cadherin, N‐cadherin and snail in shNC and shYTHDF3 in MDA‐MB‐231 and MCF‐7 cells.

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

    doi: 10.1111/jcmm.71105

    Figure Lengend Snippet: Silence of YTHDF3 restrain cell proliferation and migration of breast cancer cells and arrested the cell cycle. (A) Relative expression of YTHDF3 mRNA and protein after transfection of shNC and shYTHDF3 in MDA‐MB‐231 and MCF‐7 cells; (B) the growth ability of MDA‐MB‐231 and MCF‐7 cells transfected with shNC and shYTHDF3 was detected by CCK‐8 assay; (C) colony formation assay was used to detect the proliferation ability of MDA‐MB‐231 and MCF‐7 cells in shNC and shYTHDF3 groups; (D) the cell cycle distribution of MDA‐MB‐231 and MCF‐7 cells in shNC and shYTHDF3 groups was detected by flow cytometry; (E) expression of cyclin D1 and CDK4 in shNC and shYTHDF3 in MDA‐MB‐231 and MCF‐7 cells; (F, G) transwell invasion assay was used to detect the migration ability of breast cancer cells in the shNC group and the shYTHDF3 group in MCF‐7 and MDA‐MB‐231 cells; (H) the expression of vimentin, MMP9, E‐cadherin, N‐cadherin and snail in shNC and shYTHDF3 in MDA‐MB‐231 and MCF‐7 cells.

    Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

    Techniques: Migration, Expressing, Transfection, CCK-8 Assay, Colony Assay, Flow Cytometry, Transwell Invasion Assay

    Silencing YTHDF3 caused a decrease in lactate levels and affected glycolysis. (A) Lactate levels in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (B) extracellular acidification rate (ECAR) in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells measured by a seahorse instrument; (C) oxygen consumption rate (OCR) in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells measured by a seahorse instrument; (D) relative expression levels of hexokinase‐1 (HK1), hexokinase‐2 (HK2), PFKL, PKM, LDHA and LDHB in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (E) protein expression levels of PFKL, PKM, LDHA and LDHB in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; and (F) the correlation between YTHDF3 and PFKL, PKM, LDHA and LDHB was predicted using the GEPIA website.

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

    doi: 10.1111/jcmm.71105

    Figure Lengend Snippet: Silencing YTHDF3 caused a decrease in lactate levels and affected glycolysis. (A) Lactate levels in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (B) extracellular acidification rate (ECAR) in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells measured by a seahorse instrument; (C) oxygen consumption rate (OCR) in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells measured by a seahorse instrument; (D) relative expression levels of hexokinase‐1 (HK1), hexokinase‐2 (HK2), PFKL, PKM, LDHA and LDHB in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (E) protein expression levels of PFKL, PKM, LDHA and LDHB in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; and (F) the correlation between YTHDF3 and PFKL, PKM, LDHA and LDHB was predicted using the GEPIA website.

    Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

    Techniques: Expressing

    YTHDF3 regulates the glycolysis level of breast cancer through the mTOR–HIF1α–LDHA axis. (A) Relative mRNA expression of HIF1α in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (B) the correlation between YTHDF3 and HIF1α was predicted using the GEPIA website; (C) expression of key proteins in the mTOR–HIF1α–LDHA axis in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (D) after adding activator MHY1445, the growth ability of cancer cells in shNC and shYTHDF3 groups was detected by CCK‐8 assay; (E) the lactate level of cancer cells in shNC and shYTHDF3 groups treated with activator MHY1485; and (F) expression of proteins of cancer cells in shNC and shYTHDF3 groups treated with activator MHY1485.

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

    doi: 10.1111/jcmm.71105

    Figure Lengend Snippet: YTHDF3 regulates the glycolysis level of breast cancer through the mTOR–HIF1α–LDHA axis. (A) Relative mRNA expression of HIF1α in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (B) the correlation between YTHDF3 and HIF1α was predicted using the GEPIA website; (C) expression of key proteins in the mTOR–HIF1α–LDHA axis in shNC and shYTHDF3 groups in MDA‐MB‐231 and MCF‐7 cells; (D) after adding activator MHY1445, the growth ability of cancer cells in shNC and shYTHDF3 groups was detected by CCK‐8 assay; (E) the lactate level of cancer cells in shNC and shYTHDF3 groups treated with activator MHY1485; and (F) expression of proteins of cancer cells in shNC and shYTHDF3 groups treated with activator MHY1485.

    Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

    Techniques: Expressing, CCK-8 Assay

    YTHDF3 regulates breast cancer development through the mTOR–HIF1α–LDHA axis in vivo. (A) Tumour volumes in shNC and shYTHDF3 groups on the indicated days; (B) representative tumour images and (C) tumour weights in shNC and shYTHDF3 groups on day 63; (D) mRNA and (E) protein expression levels of YTHDF3, HIF1α and LDHA in the tumours in shNC and shYTHDF3 groups; (F) Immunohistochemical images of YTHDF3, Ki67, HIF1α and LDHA in tumours in the shNC and shYTHDF3 groups, ×100.

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

    doi: 10.1111/jcmm.71105

    Figure Lengend Snippet: YTHDF3 regulates breast cancer development through the mTOR–HIF1α–LDHA axis in vivo. (A) Tumour volumes in shNC and shYTHDF3 groups on the indicated days; (B) representative tumour images and (C) tumour weights in shNC and shYTHDF3 groups on day 63; (D) mRNA and (E) protein expression levels of YTHDF3, HIF1α and LDHA in the tumours in shNC and shYTHDF3 groups; (F) Immunohistochemical images of YTHDF3, Ki67, HIF1α and LDHA in tumours in the shNC and shYTHDF3 groups, ×100.

    Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

    Techniques: In Vivo, Expressing, Immunohistochemical staining

    YTHDF3 is associated with prognosis in several cancers. (A) Column nomogram was constructed using YTHDF3 expression levels, age, T stage, N stage, M stage and tumour stage data of breast cancer patients; (B–E) survival analysis software package was used to analyse the survival prognosis difference between patients with high and low YTHDF3 expression in BRCA, DLBC, THCA and UVM.

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: YTHDF3 Mediates the Occurrence and Development of Breast Cancer by Regulating Glycolysis Through the mTOR – HIF1α – LHDA Axis

    doi: 10.1111/jcmm.71105

    Figure Lengend Snippet: YTHDF3 is associated with prognosis in several cancers. (A) Column nomogram was constructed using YTHDF3 expression levels, age, T stage, N stage, M stage and tumour stage data of breast cancer patients; (B–E) survival analysis software package was used to analyse the survival prognosis difference between patients with high and low YTHDF3 expression in BRCA, DLBC, THCA and UVM.

    Article Snippet: Primary antibodies used in the present study were listed as follows: GAPDH (ab313650, Abcam), HIF1 alpha (ab1, Abcam), mTOR (BM4182, Boster), p‐mTOR (BM4840, Boster), RICTOR (A03195‐1, Boster), RAPTOR (BM4438, Boster), AKT (BM4390, Boster), p‐AKT (BM4744, Boster), S6K (BM4240, Boster), p‐S6K (BM4141, Boster), LDHA (A1146, ABclonal), YTHDF3 (A8395, ABclonal), cyclin D1 (A11022, ABclonal), CDK4 (A23522, ABclonal), vimentin ( PAB48967 , Bioswamp), MMP9 (FNab05247, FineTest), LDHB (A5131, ABclonal), PKM1/2 (A27199PM, ABclonal), PFKL (A23687, ABclonal), snail (13099–1‐AP, Proteintech), E‐cadherin (PB9561, Boster), N‐cadherin (A01577‐3, Boster).

    Techniques: Construct, Expressing, Software