antibody targeting ezh2  (Proteintech)

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: Elevated EZH2 expression correlates with poor prognosis in hepatocellular carcinoma (HCC). A Expression of EZH2 in the TCGA-HCC dataset, comparing HCC tissues with peri-tumoral liver tissues. B RT-qPCR assays demonstrating mRNA levels of EZH2 in 12 HCC patient samples and their corresponding adjacent normal tissues. C , D Western blot demonstrating protein levels of EZH2 in 12 HCC patient samples and their corresponding adjacent normal tissues. E Representative immunohistochemistry (IHC) images showing EZH2 expression in HCC tissues versus paired adjacent normal tissues. Statistical analysis comparing IHC scores of 12 HCC tissues with their paired normal tissues. F , G Kaplan-Meier overall survival curve and disease-free survival curve of HCC patients correlated with the expression of EZH2. All data are presented as the means ± SDs, n = 5 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: EZH2 expression is increased in lenvatinib-resistant HCC cells and contributes to resistance development. A Schematic representation of the establishment of lenvatinib-resistant (LR) cells. B Cell viability assessed by CCK-8 assay following exposure to varying concentrations of lenvatinib for 48 h. Proliferation of both Huh7-LR and Hep3B-LR cells was greater than that of their parental cells at different lenvatinib concentrations. C , D mRNA and protein levels of EZH2 in Huh7-LR and Hep3B-LR cells, as well as their parental cells, detected by RT-PCR and western blot. E , F mRNA and protein levels of EZH2 in Huh7-LR and Hep3B-LR cells transfected with shEZH2#1, shEZH2#2, or shCtrl,, detected by RT-PCR and western blot. G , H Proliferation of both Huh7-LR and Hep3B-LR cells transfected with shEZH2#1 compared to those transfected with shCtrl at varying concentrations of lenvatinib. All data are presented as the means ± SDs, n = 5 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Expressing, CCK-8 Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: EZH2 inhibits ferroptosis in HCC cells. A Analysis of cell death and apoptosis in Huh7-LR and Hep3B-LR cells after Lenvatinib (Lenv) treatment via Annexin V-FITC/PI staining. B Inhibitory ratio that was measured in Huh7-LR and Hep3B-LR using CCK-8 assay 24 h after treatment with Lenvatinib with or without Z-VAD-FMK, CQ, Nec-1 or Fer-1. C Proliferation of both Huh7-LR and Hep3B-LR cells transfected with shCtrl compared to those transfected with shEZH2#1 at varying concentrations of RSL3. D - F Relative levels of reactive oxygen species (ROS) (D), glutathione (GSH) ( E ), and malondialdehyde (MDA) ( F ) in Huh7-LR and Hep3B-LR cells transfected with shEZH2 #1 compared to shCtrl treated with Lenvatinib, RSL3 and Fer-1. (G-H) Intracellular lipid peroxidation and iron accumulation in Huh7-LR and Hep3B-LR cells transfected with shEZH2 #1 compared to shCtrl treated with Lenvatinib, RSL3 and Fer-1 were determined by C11-BODIPY and FerroOrange staining. All data are presented as the means ± SDs, n = 5 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Staining, CCK-8 Assay, Transfection

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: EZH2 suppression induced ferroptosis via enhancing ACSL1 expression through H3K27me3. A , B RT-qPCR assays demonstrated mRNA levels of indicated genes in in Huh7-LR (left panel) and Hep3B-LR (right panel) cells transfected with shEZH2 compared to shCtrl. C , D Expression levels of EZH2 in Huh7-LR ( C ) and Hep3B-LR ( D ) cells transfected with shEZH2 compared to shCtrl, detected by western blot. (E)Pearson correlation analysis of TCGA dataset revealed a significant negative correlation between EZH2 and ACSL1 expression. (F)Proliferation of Huh7-LR and Hep3B-LR cells transfected with shEZH2 or shACSL1. G - I Relative levels of ROS ( G ), GSH ( H ), and MDA ( I ) in Huh7-LR and Hep3B-LR cells transfected with shEZH2 or shACSL1. J-K Intracellular lipid peroxidation and iron accumulation in Huh7-LR and Hep3B-LR cells transfected with shEZH2 or shACSL1 were determined by C11-BODIPY and FerroOrange staining. L The protein levels of EZH2 and H3K27me3 in Huh7-LR cells transfected with shEZH2 or shCtrl were determined by western blot. M The enrichments of EZH2 and H3K27me3 on promoters of ACSL1 were determined by ChIP assays. N The altered enrichments of EZH2 and H3K27me3 on promoters of ACSL1 after EZH2 downregulatiton were determined by ChIP-qPCR assay. Normal IgG served as a negative control. All data are presented as the means ± SDs, n = 5 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Staining, ChIP-qPCR, Negative Control

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: EZH2 reduction enhances lenvatinib efficacy in treating resistant HCC in vivo. A Treatment schedule illustrating the timing of tumor inoculation and administration of treatments in BALB/c nude mice ( n = 6 per group). B , C Representative images of tumors ( B ) harvested on day 27 posttreatment, and corresponding average tumor weights ( C ) in each group. D Tumor growth curves representing the progression of Huh7-LR cell-induced tumors in mice treated with lenvatinib. E Expression levels of EZH2, H3K27me3 and ACSL1 in tumors. All data are presented as the means ± SDs, n = 6 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: In Vivo, Expressing

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: Inhibition of EZH2 enhances lenvatinib efficacy in treating hepatocellular carcinoma by promoting ACSL1-Mediated Ferroptosis via H3K27me3

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Inhibition

Journal: International immunopharmacology

Article Title: Characterization of PANoptosis-related genes with immunoregulatory features in osteoarthritis.

doi: 10.1016/j.intimp.2024.112889

Figure Lengend Snippet: Fig. 5. The correlation between key Differentially Expressed Genes Associated with Panoptosis (DE-PRGs) and immune cells. (A–G) The association of CDKN1A, EZH2, MEG3, NR4A1, PIK3R2, S100A8, and SYVN1 with immune cells.

Article Snippet: After blocking with 5 % fat-free dry milk for 60 min at room temperature, the membranes were then exposed overnight at 4 ◦C to primary antibodies targeting CDKN1A (Proteintech, China), EZH2 (Proteintech, China), NR4A1 (Proteintech, China), PIK3R2 (Proteintech, China), MEG3 (Proteintech, China), S100A8 (Proteintech, China), SYVN1 (Proteintech, China), and β-actin (1:5000).

Techniques:

Journal: International immunopharmacology

Article Title: Characterization of PANoptosis-related genes with immunoregulatory features in osteoarthritis.

doi: 10.1016/j.intimp.2024.112889

Figure Lengend Snippet: Fig. 6. Potential drug screening and primary structure of proteins. (A) CMap analysis revealed the mechanism of action associated with small-molecule compounds; (B–H) Primary protein structures of CDKN1A, EZH2, MEG3, NR4A1, PIK3R2, S100A8, and SYVN1; (I) Screening of the 3D structure of APHA-compound-8 through the PubChem open chemical database.

Article Snippet: After blocking with 5 % fat-free dry milk for 60 min at room temperature, the membranes were then exposed overnight at 4 ◦C to primary antibodies targeting CDKN1A (Proteintech, China), EZH2 (Proteintech, China), NR4A1 (Proteintech, China), PIK3R2 (Proteintech, China), MEG3 (Proteintech, China), S100A8 (Proteintech, China), SYVN1 (Proteintech, China), and β-actin (1:5000).

Techniques: Drug discovery

Journal: International immunopharmacology

Article Title: Characterization of PANoptosis-related genes with immunoregulatory features in osteoarthritis.

doi: 10.1016/j.intimp.2024.112889

Figure Lengend Snippet: Fig. 7. Molecular docking models. (A-G) Molecular docking models of CDKN1A, EZH2, MEG3, NR4A1, PIK3R2, S100A8, and SYVN1 with the potential drug (APHA- compound-8).

Article Snippet: After blocking with 5 % fat-free dry milk for 60 min at room temperature, the membranes were then exposed overnight at 4 ◦C to primary antibodies targeting CDKN1A (Proteintech, China), EZH2 (Proteintech, China), NR4A1 (Proteintech, China), PIK3R2 (Proteintech, China), MEG3 (Proteintech, China), S100A8 (Proteintech, China), SYVN1 (Proteintech, China), and β-actin (1:5000).

Techniques:

Journal: International immunopharmacology

Article Title: Characterization of PANoptosis-related genes with immunoregulatory features in osteoarthritis.

doi: 10.1016/j.intimp.2024.112889

Figure Lengend Snippet: Fig. 8. Validation by Western blot experiments. (A-H) Comparative expression analysis of CDKN1A, EZH2, MEG3, NR4A1, PIK3R2, S100A8, and SYVN1 between the normal and OA groups, all showing statistically significant differences. (***P ≤0.001).

Article Snippet: After blocking with 5 % fat-free dry milk for 60 min at room temperature, the membranes were then exposed overnight at 4 ◦C to primary antibodies targeting CDKN1A (Proteintech, China), EZH2 (Proteintech, China), NR4A1 (Proteintech, China), PIK3R2 (Proteintech, China), MEG3 (Proteintech, China), S100A8 (Proteintech, China), SYVN1 (Proteintech, China), and β-actin (1:5000).

Techniques: Biomarker Discovery, Western Blot, Expressing

Journal: bioRxiv

Article Title: Altered Huntingtin-Chromatin Interactions Predict Transcriptional and Epigenetic Changes in Huntington’s Disease

doi: 10.1101/2020.06.04.132571

Figure Lengend Snippet: A: Volcano plots summarizing differentially histone modifications for H3K27me3 and H3K4me3 and differential occupancy for EZH2 in striatal tissue from four-month-old in Htt Q111/+ vs. Htt +/+ mice. The x-axis indicates the log 2 FC and the y-axis indicates the −log 10 (p-value). Regions with greater levels or occupancy in the Htt +/+ mice are shown in orange, regions with lower levels/occupancy are indicated in blue. The dashed horizontal line indicates an FDR of 0.05, and the solid line represents a nominal p-value of 0.05. B: Enrichment of EZH2, H3K27me3 and H3K4me3 ChIP-seq peaks (MACS FDR < 0.05) at WT-specific, Q111-specific, and Shared HTT ChIP-seq peaks. Y-axis indicates the log-transformed fold change (enrichment or depletion) in the number of overlapping base pairs compared to the average from 100,000 re-sampling permutations of genomic coordinates. Plotting color indicates the p-value, derived from these same permutations. C: Gene Ontology Biological Processes enriched near differentially methylated regions with reduced H3K27me3 in Htt Q111/+ mice (GREAT ). X-axis, p-value from binomial test over genomic regions; plot color, p-value from the hypergeometric test over genes results; plot size, hypergeometric test fold-enrichment.

Article Snippet: DNA was sonicated to an average fragment length of 300-500bp, and 25ug chromatin plus 200ng Drosophila spike-in chromatin was incubated with antibody targeting EZH2, H3K27me3 or H3K4me3 (Active Motif catalog numbers 39901, 39155 and 399159, respectively).

Techniques: ChIP-sequencing, Transformation Assay, Sampling, Derivative Assay, Methylation

Journal: bioRxiv

Article Title: Altered Huntingtin-Chromatin Interactions Predict Transcriptional and Epigenetic Changes in Huntington’s Disease

doi: 10.1101/2020.06.04.132571

Figure Lengend Snippet: Each plot indicates the fold change of HTT occupancy in Htt Q111/+ / Htt +/+ mice (x-axis) compared to the fold change in the same regions for H3K27me3 (A), EZH2 (B), and H3K4me3 (C) (y-axis). Analysis is restricted to 263 HTT peaks located +/- 20 kb from the TSSs of genes that are significantly up- or down-regulated in the striatum of six-month-old knock-in mouse models of HD mutations. Each point represents a single HTT ChIP-seq peak and is labeled with the name of the adjacent differentially expressed gene. Point color indicates the fold change in gene expression in the striatum of HD knock-in mice (up = yellow; down = purple), and point size corresponds to the p-value for differential expression.

Article Snippet: DNA was sonicated to an average fragment length of 300-500bp, and 25ug chromatin plus 200ng Drosophila spike-in chromatin was incubated with antibody targeting EZH2, H3K27me3 or H3K4me3 (Active Motif catalog numbers 39901, 39155 and 399159, respectively).

Techniques: Knock-In, ChIP-sequencing, Labeling, Gene Expression, Quantitative Proteomics