Journal: EMBO Reports

Article Title: STAG2 loss in Ewing sarcoma alters enhancer-promoter contacts dependent and independent of EWS::FLI1

doi: 10.1038/s44319-024-00303-6

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Mouse monoclonal anti-EZH2 (AC22) IB: 1:1000 , CST , cat#3147.

Techniques: Recombinant, Sequencing, Reverse Transcription, SYBR Green Assay, Software, Isolation

Journal: International Journal of Molecular Sciences

Article Title: EZH2 Expression in Head-and-Neck Squamous Cell Cancer in Young Patients

doi: 10.3390/ijms25105250

Figure Lengend Snippet: Summary of the main molecular interactions (with functional outcomes italicized) of EZH2. EZH2 might act both as a PRC2 member and independently through interactions with other factors. Hypoxia, a common phenomenon in the tumor microenvironment, might up-regulate the PRC2-independent EZH2 functions both by down-regulating the expression of PRC2 members (EED and SUZ12) and by facilitating the binding of free EZH2 to FoxM1 instead of other PRC2 members, thus modulating the expression of MMPS, hence the invasive capacity of tumor cells. EZH2 itself might facilitate the dissociation of EAF2 from VHL, thus activating HIF-1α-linked transcriptional alterations, including the up-regulation of glycolitic capacity, hypoxia-tolerance, and the expression of free EZH2. The PRC2-linked effects of EZH2 also support cellular proliferation and might hinder differentiation. The tumor suppressor TP53 acts against the expression of PRC2 members (EED and EZH2) [ , , , , , , , ].

Article Snippet: Immunohistochemical staining was performed to detect EZH2 expression (mouse monoclonal anti-EZH2 antibody, clone 11, BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Functional Assay, Expressing, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: EZH2 Expression in Head-and-Neck Squamous Cell Cancer in Young Patients

doi: 10.3390/ijms25105250

Figure Lengend Snippet: ( A ) Comparison of the median proportion of EZH2-expressing cells in the two examined groups, dots represent individual data points. ( B ) comparison of the median staining intensity of EZH2 (young and control groups). Medians, interquartile ranges and total ranges are indicated.

Article Snippet: Immunohistochemical staining was performed to detect EZH2 expression (mouse monoclonal anti-EZH2 antibody, clone 11, BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Comparison, Expressing, Staining

Journal: International Journal of Molecular Sciences

Article Title: EZH2 Expression in Head-and-Neck Squamous Cell Cancer in Young Patients

doi: 10.3390/ijms25105250

Figure Lengend Snippet: Proportions of EZH2-expressing tumor cells in certain locations in the young and control HNSCC groups. Medians and interquartile ranges are indicated.

Article Snippet: Immunohistochemical staining was performed to detect EZH2 expression (mouse monoclonal anti-EZH2 antibody, clone 11, BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: EZH2 Expression in Head-and-Neck Squamous Cell Cancer in Young Patients

doi: 10.3390/ijms25105250

Figure Lengend Snippet: Staining characteristics of tumor cells of the young patient and control HNSCC groups.

Article Snippet: Immunohistochemical staining was performed to detect EZH2 expression (mouse monoclonal anti-EZH2 antibody, clone 11, BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Staining, Expressing, Comparison

Journal: International Journal of Molecular Sciences

Article Title: EZH2 Expression in Head-and-Neck Squamous Cell Cancer in Young Patients

doi: 10.3390/ijms25105250

Figure Lengend Snippet: Cox analysis for the predictors of patient survival among young HNSCC-afflicted individuals.

Article Snippet: Immunohistochemical staining was performed to detect EZH2 expression (mouse monoclonal anti-EZH2 antibody, clone 11, BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: EZH2 Expression in Head-and-Neck Squamous Cell Cancer in Young Patients

doi: 10.3390/ijms25105250

Figure Lengend Snippet: Representative samples and two different degrees of EZH2 expression (proportions of positive nuclei: ( A ) 20%; ( B ) 80%; ( C ) 100%). Tumor tissue can be seen in the submucosa; the dark tone in the nucleus indicates EZH2 expression. Positivity in the basal layer of physiological mucosal epithelium is normal. In the pictures, EZH2 expression extends to the full length of the epithelium due to the in situ carcinoma or epithelial dysplasia in the tumor environment. Scale bars represent 50 μm, on picture ( A ), 100 μm, on picture ( B ), 200 μm, on picture ( C ).

Article Snippet: Immunohistochemical staining was performed to detect EZH2 expression (mouse monoclonal anti-EZH2 antibody, clone 11, BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Expressing, In Situ

Journal: Frontiers in Cell and Developmental Biology

Article Title: Dual inhibition of EZH2 and G9A/GLP histone methyltransferases by HKMTI-1-005 promotes differentiation of acute myeloid leukemia cells

doi: 10.3389/fcell.2023.1076458

Figure Lengend Snippet: Knockdown of EZH2 promotes differentiation of HL-60 AML cells by ATRA. (A) Expression of the myeloid differentiation marker CD11b (left panel), and proportion of live cells (PI-negative population, right panel) following lentivirally-mediated knockdown (KD) of EZH2 expression and analysis by flow cytometry. EZH2 KD was achieved using two different short-hairpin RNA sequences (shRNA #1 and #2) as indicated. A non-targeting shRNA was used as a negative control. Values represent the means of three experiments and error bars denote standard deviations. * p < 0.05; *** p < 0.001; ns, no statistical significance. (B) Immunoblot analysis of levels of EZH2 protein and trimethylated H3 Lys27 (H3K27 me3 ) following EZH2 KD. Transduced cells were selected with puromycin and expanded for a minimum of 10 days. GAPDH was used as a loading control. (C) Cell morphology of cells analyzed by May-Grunwald Giemsa staining following EZH2 KD (shRNA #1) +/− ATRA (0.1 µM for 72 h).

Article Snippet: Antibodies used for immunoblot and immunoprecipitation were as follows: rabbit polyclonal anti-H3K27 me3 (Cat# 07–473, Upstate Biotechnology); rabbit polyclonal anti-H3K9 me2 (Cat# ab115159, Abcam); rabbit polyclonal anti-GAPDH (Cat# ab9485, Abcam); rabbit polyclonal anti-EZH2 (Cat# ab186006, Abcam); mouse monoclonal anti-EZH2 (clone AC22, Cat# 3147, Cell Signaling Technology); mouse monoclonal anti-RARα (clone 9α-9A6, Cat# 39971, Active Motif); rabbit polyclonal anti-RARα (Cat# sc-773, Santa Cruz).

Techniques: Expressing, Marker, Flow Cytometry, shRNA, Negative Control, Western Blot, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Dual inhibition of EZH2 and G9A/GLP histone methyltransferases by HKMTI-1-005 promotes differentiation of acute myeloid leukemia cells

doi: 10.3389/fcell.2023.1076458

Figure Lengend Snippet: Dual inhibition of EZH2-G9A/GLP by HKMTI-1-005 promotes differentiation of HL-60 AML cells by ATRA. (A) Proportion of viable cells as determined by CellTiter-Glo cell viability assay (left panel), and expression of the myeloid differentiation marker CD11b (right panel) following treatment with the indicated concentrations of GSK-343 +/− 0.1 µM ATRA for 72 h. (B) Proportion of viable cells as determined by CellTiter-Glo cell viability assay (left panel) and expression of the myeloid differentiation marker CD11b (right panel) following treatment with the indicated concentrations of HKMTI-1-005 +/− 0.1 µM ATRA for 72 h. Values represent the means of three experiments (viability assays) or two experiments (CD11b flow cytometry). Error bars denote standard deviations. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns, no statistical significance. (C) Cell morphology of cells analyzed by May-Grunwald Giemsa staining following treatment with 10 µM GSK-343 (left panel) or 2.5 µM HKMTI-1-005 (right panel) for 72 h. Treatments were performed +/− 0.1 µM ATRA. (D) Immunoblot analysis of levels of trimethylated H3 Lys27 (H3K27 me3 ), dimethylated H3 Lys9 (H3K9 me2 ) and trimethylated H3 Lys9 (H3K9 me3 ) following treatment (72 h) with 10 µM GSK-343 or 2.5 µM HKMTI-1-005 +/− 0.1 µM ATRA as indicated. GAPDH was used as a loading control.

Article Snippet: Antibodies used for immunoblot and immunoprecipitation were as follows: rabbit polyclonal anti-H3K27 me3 (Cat# 07–473, Upstate Biotechnology); rabbit polyclonal anti-H3K9 me2 (Cat# ab115159, Abcam); rabbit polyclonal anti-GAPDH (Cat# ab9485, Abcam); rabbit polyclonal anti-EZH2 (Cat# ab186006, Abcam); mouse monoclonal anti-EZH2 (clone AC22, Cat# 3147, Cell Signaling Technology); mouse monoclonal anti-RARα (clone 9α-9A6, Cat# 39971, Active Motif); rabbit polyclonal anti-RARα (Cat# sc-773, Santa Cruz).

Techniques: Inhibition, Viability Assay, Expressing, Marker, Flow Cytometry, Staining, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Dual inhibition of EZH2 and G9A/GLP histone methyltransferases by HKMTI-1-005 promotes differentiation of acute myeloid leukemia cells

doi: 10.3389/fcell.2023.1076458

Figure Lengend Snippet: Differential gene expression following EZH2 knockdown or inhibition with GSK-343 or HKMTI-1-005 in HL-60 AML cells. (A) Principal component analysis (PCA; covariance) of GCRMA-normalized expression array samples in biological triplicates represented with centroids as annotated: two different EZH2 -targeting shRNAs and one non-targeting control (days ∼10–20 post transduction), 10 µM GSK-343 or 2.5 µM HKMTI-1-005 +/− 0.1 µM ATRA following treatment for 72 h. Red or blue ellipses highlight specific grouping of samples of interest. (B) Gene ontology (GO) analysis of upregulated genes following EZH2 KD or drug treatment as annotated (0.1 µM ATRA, 10 µM GSK-343, 2.5 µM HKMTI-1-005 for 72 h) showing enrichment score (left panel) and fold-enrichment over control (right panel) of genes belonging to the indicated categories. (C) Venn diagram showing groups of genes taken for further analysis that belong exclusively to GSK-343 treated cells, or are shared only between EZH2 KD and HKMTI-1-005 treated cells, or between EZH2 KD, HKMTI-1-005 and ATRA, but not GSK-343 treated cells. Gene groups are shaded grey and indicated.

Article Snippet: Antibodies used for immunoblot and immunoprecipitation were as follows: rabbit polyclonal anti-H3K27 me3 (Cat# 07–473, Upstate Biotechnology); rabbit polyclonal anti-H3K9 me2 (Cat# ab115159, Abcam); rabbit polyclonal anti-GAPDH (Cat# ab9485, Abcam); rabbit polyclonal anti-EZH2 (Cat# ab186006, Abcam); mouse monoclonal anti-EZH2 (clone AC22, Cat# 3147, Cell Signaling Technology); mouse monoclonal anti-RARα (clone 9α-9A6, Cat# 39971, Active Motif); rabbit polyclonal anti-RARα (Cat# sc-773, Santa Cruz).

Techniques: Expressing, Inhibition, Transduction

Journal: Frontiers in Cell and Developmental Biology

Article Title: Dual inhibition of EZH2 and G9A/GLP histone methyltransferases by HKMTI-1-005 promotes differentiation of acute myeloid leukemia cells

doi: 10.3389/fcell.2023.1076458

Figure Lengend Snippet: Gene set enrichment analysis (GSEA) of gene expression in HL-60 cells reveals opposite effects of GSK-343 and HKMTI-1-005 on stem cell and differentiation programs. Gene expression data were generated by expression microarray analysis following 72 h treatment with 10 µM GSK-343 or 2.5 µM HKMTI-1-005, or EZH2 KD versus untreated control and analyzed using GSEA to extract biological knowledge. Highly significantly enriched gene sets are indicated by their standard and systematic names. The most upregulated genes in treatment or EZH2 KD samples are shown on the left side (red), while the most upregulated genes in the control are shown on the right side (blue). Black bars represent the positions of the treatment or EZH2 KD samples versus vehicle control upregulated signature genes in the ranked list. Green curves represent the evolution of gene density. Normalized enrichment scores (NES) reflect the degree to which genes were overrepresented. When the distribution is random, the enrichment score is zero. Enrichment of signature genes at the top of the ranked list results in a large positive deviation of the NES from zero. q -value = FDR-adjusted p -value.

Article Snippet: Antibodies used for immunoblot and immunoprecipitation were as follows: rabbit polyclonal anti-H3K27 me3 (Cat# 07–473, Upstate Biotechnology); rabbit polyclonal anti-H3K9 me2 (Cat# ab115159, Abcam); rabbit polyclonal anti-GAPDH (Cat# ab9485, Abcam); rabbit polyclonal anti-EZH2 (Cat# ab186006, Abcam); mouse monoclonal anti-EZH2 (clone AC22, Cat# 3147, Cell Signaling Technology); mouse monoclonal anti-RARα (clone 9α-9A6, Cat# 39971, Active Motif); rabbit polyclonal anti-RARα (Cat# sc-773, Santa Cruz).

Techniques: Expressing, Generated, Microarray

Journal: Frontiers in Cell and Developmental Biology

Article Title: Dual inhibition of EZH2 and G9A/GLP histone methyltransferases by HKMTI-1-005 promotes differentiation of acute myeloid leukemia cells

doi: 10.3389/fcell.2023.1076458

Figure Lengend Snippet: ATRA-mediated differentiation promotes an association between RARα and EZH2 in HL-60 cells that is inhibited by GSK-343 and enhanced by HKMTI-1-005. (A) Proximity ligation assay (PLA) analyzing RARα/EZH2 complexes following 0.1 µM ATRA treatment for the indicated times. (B) PLA analyzing RARα/EZH2 complexes following treatment with 10 µM GSK-343 or 2.5 µM HKMTI-1-005 for 4 h. Treatments were performed +/− 0.1 µM ATRA as indicated. Upper panels show representative fields of view of cells indicating proximity (< 40 nm) of antibody conjugated PLA probes that have been ligated, amplified, and detected with complementary fluorescent probes. Red dots represent the presence of RARα/EZH2 interactions. Scale bars, 20 µm. Cell nuclei are counterstained with DAPI (blue). Lower panels show mean values of signals (red dots) per cell representing RARα/EZH2 interactions. Values represent the means of two experiments (A) or three experiments (B) , and error bars denote standard deviations. ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns, no statistical significance.

Article Snippet: Antibodies used for immunoblot and immunoprecipitation were as follows: rabbit polyclonal anti-H3K27 me3 (Cat# 07–473, Upstate Biotechnology); rabbit polyclonal anti-H3K9 me2 (Cat# ab115159, Abcam); rabbit polyclonal anti-GAPDH (Cat# ab9485, Abcam); rabbit polyclonal anti-EZH2 (Cat# ab186006, Abcam); mouse monoclonal anti-EZH2 (clone AC22, Cat# 3147, Cell Signaling Technology); mouse monoclonal anti-RARα (clone 9α-9A6, Cat# 39971, Active Motif); rabbit polyclonal anti-RARα (Cat# sc-773, Santa Cruz).

Techniques: Proximity Ligation Assay, Amplification

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: STEAP and EZH2 expression in LC specimens . (A) STEAP, adenocarcinoma (×400). (B) STEAP, squamous cell carcinoma (×400). (C) EZH2, adenocarcinoma (×200). (D) EZH2, squamous cell carcinoma (×200).

Article Snippet: To detect EZH2, monoclonal mouse anti-human EZH2 (BD.612666, BD Bioscience, San Jose, CA, USA), diluted 1:200, served as the primary antibody.

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Expression of STEAP and EZH2 protein in various tumor cell lines . Western blotting was done using STEAP- or EZH2-specific antibody as described in "Materials and methods". The STEAP protein has a mass of approximately 36 kDa, and EZH2 protein 91 kDa. (A) STEAP, (B) EZH2.

Article Snippet: To detect EZH2, monoclonal mouse anti-human EZH2 (BD.612666, BD Bioscience, San Jose, CA, USA), diluted 1:200, served as the primary antibody.

Techniques: Expressing, Western Blot

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: HLA-DR restriction analysis of STEAP 281-296 - and EZH2 95-109 -reactive CD4 T-cell clones . (A) L243 but not W6/32 inhibited IFN-γ production from HLA-DR restricted, STEAP 281-296 -reactive CD4 T-cell clones. (B) HLA-DR restriction analysis of EZH2 95-109 -reactive CD4 T-cell clones. (C and D) STEAP 281-296 - and EZH2 95-109 -reactive CD4 T-cell clones were tested for their capacity to recognize autologous PBMCs as APCs in the presence of various concentrations of peptide. Points means of triplicate determinations, bars SD. Points without bars had SD < 10% the value of the mean. Results are representative of two experiments that were done with the same samples.

Article Snippet: To detect EZH2, monoclonal mouse anti-human EZH2 (BD.612666, BD Bioscience, San Jose, CA, USA), diluted 1:200, served as the primary antibody.

Techniques: Clone Assay

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Definition of restricting HLA-DR alleles of the STEAP- or EZH2-reactive CD4 T-cell clones . IFN-γ production of the STEAP 281-296 -reactive CD4 T-cell clones (A) and EZH2 95-109 -reactive CD4 T-cell clones (B, C, and D) was evaluated using L-cells or semi-allogenic EBV-LCLs homozygous for HLA-DR molecules as APCs to define the restricting HLA-DR elements. Columns means of triplicate determinations, bars SD. Columns without bars had SD < 10% the values of the mean. Results are representative of at least two experiments.

Article Snippet: To detect EZH2, monoclonal mouse anti-human EZH2 (BD.612666, BD Bioscience, San Jose, CA, USA), diluted 1:200, served as the primary antibody.

Techniques: Clone Assay

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Granzyme B production from STEAP 281-296 - or EZH2 95-109 -reactive CD4 T-cell clones . STEAP 281-296 -reactive, DR15-restricted CD4 T-cell clone SH31 (A, left) and DR53-restricted CD4 T-cell clone Sa12 (A, right) were able to secrete granzyme B reacting with STEAP+/HLA-DR matched LC cells. EZH2 95-109 -reactive, DR15-restricted CD4 T-cell clone SK32 (B, left) and Sa361 (B, middle), and DR53-restricted CD4 T-cell clone Sa362 (B, right) were able to secrete granzyme B reacting with EZH2+/HLA-DR matched LC cells. The HLA-DR-negative Jurkat was also used as negative controls. The antigen specificity of these responses was demonstrated by blocking tumor recognition with L243. Columns without bars had SD < 10% the values of the mean. Results are representative of two separate experiments.

Article Snippet: To detect EZH2, monoclonal mouse anti-human EZH2 (BD.612666, BD Bioscience, San Jose, CA, USA), diluted 1:200, served as the primary antibody.

Techniques: Clone Assay, Blocking Assay

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Direct recognition of EZH2-expressing lung tumor cells by EZH2 95-109 -reactive CD4 T-cell clones . DR1-restricted CD4 T-cell clone SK31 (A), DR15-restricted CD4 T-cell clone SK32 (B), Sa362 (C), DR53-restricted CD4 T-cell clones Sa362 (D) and YM18 (E) were tested for their capacity to recognize antigen directly on EZH2 positive LC cells that are either matched or mismatched for the restricting HLA-DR alleles. The HLA-DR-negative Jurkat was also used as negative controls. The antigen specificity of these responses was demonstrated by blocking tumor recognition with L243. Columns without bars had SD < 10% the values of the mean. Results are representative of two separate experiments.

Article Snippet: To detect EZH2, monoclonal mouse anti-human EZH2 (BD.612666, BD Bioscience, San Jose, CA, USA), diluted 1:200, served as the primary antibody.

Techniques: Expressing, Clone Assay, Blocking Assay

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Assessment of cytolytic activity of STEAP 281-296 - or EZH2 95-109 -reactive CD4 T-cell clones . The DR15-restricted, STEAP 281-296 -reactive CD4 T-cell clone SH31 (A, left), and DR53-restricted, STEAP 281-296 -reactive CD4 T-cell clone Sa12 (A, right) were evaluated for their capacity to kill STEAP-expressing, DR53 positive LC cells (LU65 and Calu-1) or STEAP-expressing, DR53 negative LC cells (EBC1, negative control). The DR15-restricted, EZH2 95-109 -reactive CD4 T-cell clones SK32 (B, left), Sa361 (B, middle), and DR53-restricted, EZH2 95-109 -reactive CD4 T-cell clone Sa362 (B, right) were evaluated for their capacity to kill EZH2-expressing, DR15 positive LC cells (LU65 and RERF-LC-AI), DR53 positive LC cells (LU65 and Calu-1) or EZH2-expressing, DR15 and DR53 negative LC cells (EBC1, negative control). Points means of triplicate determinations, bars SD. Points without bars had SD < 10% the value of the mean.

Article Snippet: To detect EZH2, monoclonal mouse anti-human EZH2 (BD.612666, BD Bioscience, San Jose, CA, USA), diluted 1:200, served as the primary antibody.

Techniques: Activity Assay, Clone Assay, Expressing, Negative Control

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Assessment of T-cell responses to the STEAP 281-296 and EZH2 95-109 -peptides in lung cancer patients and healthy donors.

Article Snippet: To detect EZH2, monoclonal mouse anti-human EZH2 (BD.612666, BD Bioscience, San Jose, CA, USA), diluted 1:200, served as the primary antibody.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: STEAP and EZH2 expression in LC specimens . (A) STEAP, adenocarcinoma (×400). (B) STEAP, squamous cell carcinoma (×400). (C) EZH2, adenocarcinoma (×200). (D) EZH2, squamous cell carcinoma (×200).

Article Snippet: The membrane was blocked in PBS containing 0.01% Tween 20 and 5% non-fat dry milk for 1 h at room temperature and incubated with rabbit anti-human STEAP polyclonal antibody (H-105, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:200 in blocking buffer, anti-human EZH2 mouse monoclonal antibody (mAb) (BD.612666, BD Bioscience, San Jose, CA, USA) diluted 1:200, or anti-β-actin mAb (C4, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:1,000 in blocking buffer as the control, for 18 h at 4°C.

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Expression of STEAP and EZH2 protein in various tumor cell lines . Western blotting was done using STEAP- or EZH2-specific antibody as described in "Materials and methods". The STEAP protein has a mass of approximately 36 kDa, and EZH2 protein 91 kDa. (A) STEAP, (B) EZH2.

Article Snippet: The membrane was blocked in PBS containing 0.01% Tween 20 and 5% non-fat dry milk for 1 h at room temperature and incubated with rabbit anti-human STEAP polyclonal antibody (H-105, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:200 in blocking buffer, anti-human EZH2 mouse monoclonal antibody (mAb) (BD.612666, BD Bioscience, San Jose, CA, USA) diluted 1:200, or anti-β-actin mAb (C4, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:1,000 in blocking buffer as the control, for 18 h at 4°C.

Techniques: Expressing, Western Blot

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: HLA-DR restriction analysis of STEAP 281-296 - and EZH2 95-109 -reactive CD4 T-cell clones . (A) L243 but not W6/32 inhibited IFN-γ production from HLA-DR restricted, STEAP 281-296 -reactive CD4 T-cell clones. (B) HLA-DR restriction analysis of EZH2 95-109 -reactive CD4 T-cell clones. (C and D) STEAP 281-296 - and EZH2 95-109 -reactive CD4 T-cell clones were tested for their capacity to recognize autologous PBMCs as APCs in the presence of various concentrations of peptide. Points means of triplicate determinations, bars SD. Points without bars had SD < 10% the value of the mean. Results are representative of two experiments that were done with the same samples.

Article Snippet: The membrane was blocked in PBS containing 0.01% Tween 20 and 5% non-fat dry milk for 1 h at room temperature and incubated with rabbit anti-human STEAP polyclonal antibody (H-105, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:200 in blocking buffer, anti-human EZH2 mouse monoclonal antibody (mAb) (BD.612666, BD Bioscience, San Jose, CA, USA) diluted 1:200, or anti-β-actin mAb (C4, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:1,000 in blocking buffer as the control, for 18 h at 4°C.

Techniques: Clone Assay

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Definition of restricting HLA-DR alleles of the STEAP- or EZH2-reactive CD4 T-cell clones . IFN-γ production of the STEAP 281-296 -reactive CD4 T-cell clones (A) and EZH2 95-109 -reactive CD4 T-cell clones (B, C, and D) was evaluated using L-cells or semi-allogenic EBV-LCLs homozygous for HLA-DR molecules as APCs to define the restricting HLA-DR elements. Columns means of triplicate determinations, bars SD. Columns without bars had SD < 10% the values of the mean. Results are representative of at least two experiments.

Article Snippet: The membrane was blocked in PBS containing 0.01% Tween 20 and 5% non-fat dry milk for 1 h at room temperature and incubated with rabbit anti-human STEAP polyclonal antibody (H-105, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:200 in blocking buffer, anti-human EZH2 mouse monoclonal antibody (mAb) (BD.612666, BD Bioscience, San Jose, CA, USA) diluted 1:200, or anti-β-actin mAb (C4, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:1,000 in blocking buffer as the control, for 18 h at 4°C.

Techniques: Clone Assay

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Granzyme B production from STEAP 281-296 - or EZH2 95-109 -reactive CD4 T-cell clones . STEAP 281-296 -reactive, DR15-restricted CD4 T-cell clone SH31 (A, left) and DR53-restricted CD4 T-cell clone Sa12 (A, right) were able to secrete granzyme B reacting with STEAP+/HLA-DR matched LC cells. EZH2 95-109 -reactive, DR15-restricted CD4 T-cell clone SK32 (B, left) and Sa361 (B, middle), and DR53-restricted CD4 T-cell clone Sa362 (B, right) were able to secrete granzyme B reacting with EZH2+/HLA-DR matched LC cells. The HLA-DR-negative Jurkat was also used as negative controls. The antigen specificity of these responses was demonstrated by blocking tumor recognition with L243. Columns without bars had SD < 10% the values of the mean. Results are representative of two separate experiments.

Article Snippet: The membrane was blocked in PBS containing 0.01% Tween 20 and 5% non-fat dry milk for 1 h at room temperature and incubated with rabbit anti-human STEAP polyclonal antibody (H-105, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:200 in blocking buffer, anti-human EZH2 mouse monoclonal antibody (mAb) (BD.612666, BD Bioscience, San Jose, CA, USA) diluted 1:200, or anti-β-actin mAb (C4, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:1,000 in blocking buffer as the control, for 18 h at 4°C.

Techniques: Clone Assay, Blocking Assay

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Direct recognition of EZH2-expressing lung tumor cells by EZH2 95-109 -reactive CD4 T-cell clones . DR1-restricted CD4 T-cell clone SK31 (A), DR15-restricted CD4 T-cell clone SK32 (B), Sa362 (C), DR53-restricted CD4 T-cell clones Sa362 (D) and YM18 (E) were tested for their capacity to recognize antigen directly on EZH2 positive LC cells that are either matched or mismatched for the restricting HLA-DR alleles. The HLA-DR-negative Jurkat was also used as negative controls. The antigen specificity of these responses was demonstrated by blocking tumor recognition with L243. Columns without bars had SD < 10% the values of the mean. Results are representative of two separate experiments.

Article Snippet: The membrane was blocked in PBS containing 0.01% Tween 20 and 5% non-fat dry milk for 1 h at room temperature and incubated with rabbit anti-human STEAP polyclonal antibody (H-105, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:200 in blocking buffer, anti-human EZH2 mouse monoclonal antibody (mAb) (BD.612666, BD Bioscience, San Jose, CA, USA) diluted 1:200, or anti-β-actin mAb (C4, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:1,000 in blocking buffer as the control, for 18 h at 4°C.

Techniques: Expressing, Clone Assay, Blocking Assay

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Assessment of cytolytic activity of STEAP 281-296 - or EZH2 95-109 -reactive CD4 T-cell clones . The DR15-restricted, STEAP 281-296 -reactive CD4 T-cell clone SH31 (A, left), and DR53-restricted, STEAP 281-296 -reactive CD4 T-cell clone Sa12 (A, right) were evaluated for their capacity to kill STEAP-expressing, DR53 positive LC cells (LU65 and Calu-1) or STEAP-expressing, DR53 negative LC cells (EBC1, negative control). The DR15-restricted, EZH2 95-109 -reactive CD4 T-cell clones SK32 (B, left), Sa361 (B, middle), and DR53-restricted, EZH2 95-109 -reactive CD4 T-cell clone Sa362 (B, right) were evaluated for their capacity to kill EZH2-expressing, DR15 positive LC cells (LU65 and RERF-LC-AI), DR53 positive LC cells (LU65 and Calu-1) or EZH2-expressing, DR15 and DR53 negative LC cells (EBC1, negative control). Points means of triplicate determinations, bars SD. Points without bars had SD < 10% the value of the mean.

Article Snippet: The membrane was blocked in PBS containing 0.01% Tween 20 and 5% non-fat dry milk for 1 h at room temperature and incubated with rabbit anti-human STEAP polyclonal antibody (H-105, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:200 in blocking buffer, anti-human EZH2 mouse monoclonal antibody (mAb) (BD.612666, BD Bioscience, San Jose, CA, USA) diluted 1:200, or anti-β-actin mAb (C4, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:1,000 in blocking buffer as the control, for 18 h at 4°C.

Techniques: Activity Assay, Clone Assay, Expressing, Negative Control

Journal: Journal of Translational Medicine

Article Title: Six-transmembrane epithelial antigen of the prostate and enhancer of zeste homolog 2 as immunotherapeutic targets for lung cancer

doi: 10.1186/1479-5876-9-191

Figure Lengend Snippet: Assessment of T-cell responses to the STEAP 281-296 and EZH2 95-109 -peptides in lung cancer patients and healthy donors.

Article Snippet: The membrane was blocked in PBS containing 0.01% Tween 20 and 5% non-fat dry milk for 1 h at room temperature and incubated with rabbit anti-human STEAP polyclonal antibody (H-105, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:200 in blocking buffer, anti-human EZH2 mouse monoclonal antibody (mAb) (BD.612666, BD Bioscience, San Jose, CA, USA) diluted 1:200, or anti-β-actin mAb (C4, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:1,000 in blocking buffer as the control, for 18 h at 4°C.

Techniques:

Journal: BMC Genomics

Article Title: Identification of microRNA-mRNA functional interactions in UVB-induced senescence of human diploid fibroblasts

doi: 10.1186/1471-2164-14-224

Figure Lengend Snippet: Transcriptome analysis of UVB-induced senescence. A . Ingenuity pathway analysis. Activated pathways were determined by Ingenuity System Pathway Analysis software ( http://www.ingenuity.com/ ) by Core Analysis. Shown are eight significantly activated pathways (threshold p-value ≤ 0.05). B . In silico analysis of mRNA expression change in response to UVB irradiation. (left) The differential gene expression profile from total RNA was examined by Affymetrix GeneChip analysis (cutoff of > 1.5 fold regulation) 1, 7, and 9 days between control and UVB irradiated of HDF samples. The expression values were sorted by level of Ezh2 expression (rows). (right) Heatmap of selected 67 genes were determined by RT-PCR and sorted by level of Ezh2 expression (rows), red representing overexpression and green representing underexpression of the transcript.

Article Snippet: The following antibodies were used: mouse monoclonal anti-p21 WAF1 (Pharmingen, #556430) mouse monoclonal anti-pRb (Pharmingen, #554136), mouse monoclonal anti-p53 (Santa Cruz, #sc-126), rabbit polyclonal anti-GAPDH (Santa Cruz, #sc-25778), rabbit polyclonal phospho-p53 (Ser15; Cell Signaling, #9284), mouse monoclonal α-Tubulin (Sigma, #t-5168), mouse monoclonal ant-Cyclin D1 (Neomarkers, #MS-210-P) and mouse monoclonal anti-Ezh2 (BD Biosciences, #612666), mouse monoclonal anti-p16 INK4A (BD Biosciences, #511325GR), rabbit polyclonal anti-p15 INK4b (Abcam, #ab53034), mouse monoclonal anti-RUNX1/AML1 (Abcam, #ab54869), rabbit monoclonal anti-retinoic acid receptor beta – RARB (abcam, #ab124701).

Techniques: Software, In Silico, Expressing, Irradiation, Reverse Transcription Polymerase Chain Reaction, Over Expression

Journal: BMC Genomics

Article Title: Identification of microRNA-mRNA functional interactions in UVB-induced senescence of human diploid fibroblasts

doi: 10.1186/1471-2164-14-224

Figure Lengend Snippet: Correlation network of miR-101 and its high confidence target genes. (A) We used 10 prediction tools to obtain, based on public data, candidate miRNA-mRNA target interactions and we identified high confidence targets by mRNA and miRNA expression. We calculated Pearson correlation coefficients between miRNAs and their targets. Results of the analysis are presented here for miR-101. The color and shape of nodes are based on different node attributes available for the analyzed dataset. The red triangles, purple circles and orange diamonds in the network are indicating miRNAs, target genes, and transcription factors, respectively. Edges represent correlation between miRNAs and mRNAs, the color of the edges designate the type of interaction. Red is for positive and green is for negative correlation. (B) Protein samples for the indicated time points were collected as described (left panel). Protein levels were determined for Ezh2 by standard Western blot analysis. Experiments were performed in triplicates, shown here is a representative result. miR-101 levels were determined by real-time qPCR as described (right panel). Data represents the mean ± SE for three independent experiments. Co: untreated controls. *p < 0.01; **p < 0.001.

Article Snippet: The following antibodies were used: mouse monoclonal anti-p21 WAF1 (Pharmingen, #556430) mouse monoclonal anti-pRb (Pharmingen, #554136), mouse monoclonal anti-p53 (Santa Cruz, #sc-126), rabbit polyclonal anti-GAPDH (Santa Cruz, #sc-25778), rabbit polyclonal phospho-p53 (Ser15; Cell Signaling, #9284), mouse monoclonal α-Tubulin (Sigma, #t-5168), mouse monoclonal ant-Cyclin D1 (Neomarkers, #MS-210-P) and mouse monoclonal anti-Ezh2 (BD Biosciences, #612666), mouse monoclonal anti-p16 INK4A (BD Biosciences, #511325GR), rabbit polyclonal anti-p15 INK4b (Abcam, #ab53034), mouse monoclonal anti-RUNX1/AML1 (Abcam, #ab54869), rabbit monoclonal anti-retinoic acid receptor beta – RARB (abcam, #ab124701).

Techniques: Expressing, Western Blot

Journal: BMC Genomics

Article Title: Identification of microRNA-mRNA functional interactions in UVB-induced senescence of human diploid fibroblasts

doi: 10.1186/1471-2164-14-224

Figure Lengend Snippet: Depletion of miR-101 fails to prevent UVB-induced senescence. Cells were irradiated with UVB twice a day for 4 days. After the last exposure the cells were reverse transfected as described with miR-101 power inhibitors for knockdown, negative control or not treated. Reverse transfection was repeated at day 7. At day 9 RNA and protein were isolated. A . Control cells. Expression levels of miR-101 and Ezh2 mRNA of non-irradiated cells after miR-101 knockdown were determined by real-time qPCR. Bars indicate the mean ± SE of three independent experiments. B . UVB-treated cells miR-101 and Ezh2 expression levels from UVB treated cells after miR-101 knockdown were determined by real-time qPCR. Bars indicate the mean ± SE of three independent experiments. C . Growth curve analysis of miR-101 knockdown cells. cPDLs were calculated as described. Data represent the mean ± SD of three independent experiments. cPDL: cumulative population doublings KD: knockdown. *p < 0.01; **p < 0.001.

Article Snippet: The following antibodies were used: mouse monoclonal anti-p21 WAF1 (Pharmingen, #556430) mouse monoclonal anti-pRb (Pharmingen, #554136), mouse monoclonal anti-p53 (Santa Cruz, #sc-126), rabbit polyclonal anti-GAPDH (Santa Cruz, #sc-25778), rabbit polyclonal phospho-p53 (Ser15; Cell Signaling, #9284), mouse monoclonal α-Tubulin (Sigma, #t-5168), mouse monoclonal ant-Cyclin D1 (Neomarkers, #MS-210-P) and mouse monoclonal anti-Ezh2 (BD Biosciences, #612666), mouse monoclonal anti-p16 INK4A (BD Biosciences, #511325GR), rabbit polyclonal anti-p15 INK4b (Abcam, #ab53034), mouse monoclonal anti-RUNX1/AML1 (Abcam, #ab54869), rabbit monoclonal anti-retinoic acid receptor beta – RARB (abcam, #ab124701).

Techniques: Irradiation, Transfection, Negative Control, Isolation, Expressing

Journal: Journal of Translational Medicine

Article Title: Poorly differentiated synovial sarcoma is associated with high expression of enhancer of zeste homologue 2 (EZH2)

doi: 10.1186/1479-5876-10-216

Figure Lengend Snippet: Photomicrographs representing the scores 0, 3, and 6 for EZH2, H3K27me3, and Ki-67 immunostaning.

Article Snippet: Monoclonal mouse anti-EZH2 (1:25, clone 11, BD Biosciences, USA), monoclonal rabbit anti-trimethyl-Histone H3 Lys27 (1:200, clone C36B11, Cell Signaling Technology, USA), or monoclonal mouse anti-Ki-67 (1:50, clone MIB-1; Dako, Denmark) antibodies were applied on the slides.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Poorly differentiated synovial sarcoma is associated with high expression of enhancer of zeste homologue 2 (EZH2)

doi: 10.1186/1479-5876-10-216

Figure Lengend Snippet: Distribution of scores 0–6 in the MPSS, BPSS, and PDSS subtypes, shown as stacked columns (A). mRNA expression of EZH2 in the PDSS vs. MPSS+BPSS groups, mean ± S.D. Human ribosomal protein S18 was used as endogenous control and human lymph node was served as calibrator.

Article Snippet: Monoclonal mouse anti-EZH2 (1:25, clone 11, BD Biosciences, USA), monoclonal rabbit anti-trimethyl-Histone H3 Lys27 (1:200, clone C36B11, Cell Signaling Technology, USA), or monoclonal mouse anti-Ki-67 (1:50, clone MIB-1; Dako, Denmark) antibodies were applied on the slides.

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: Poorly differentiated synovial sarcoma is associated with high expression of enhancer of zeste homologue 2 (EZH2)

doi: 10.1186/1479-5876-10-216

Figure Lengend Snippet: Comparison of immunohistochemical scores across patient groups sorted by histological subtype or clinical / molecular characteristics

Article Snippet: Monoclonal mouse anti-EZH2 (1:25, clone 11, BD Biosciences, USA), monoclonal rabbit anti-trimethyl-Histone H3 Lys27 (1:200, clone C36B11, Cell Signaling Technology, USA), or monoclonal mouse anti-Ki-67 (1:50, clone MIB-1; Dako, Denmark) antibodies were applied on the slides.

Techniques: Immunohistochemical staining

Journal: Journal of Translational Medicine

Article Title: Poorly differentiated synovial sarcoma is associated with high expression of enhancer of zeste homologue 2 (EZH2)

doi: 10.1186/1479-5876-10-216

Figure Lengend Snippet: Scatter graphs with linear trend lines indicate positive correlations between EZH2 and the other markers. All patients were included in (A) and (B) , while PDSS patients were excluded in (C) . Spearman’s r (ρ) and coefficient of determination (R 2 ) values are shown at the bottom.

Article Snippet: Monoclonal mouse anti-EZH2 (1:25, clone 11, BD Biosciences, USA), monoclonal rabbit anti-trimethyl-Histone H3 Lys27 (1:200, clone C36B11, Cell Signaling Technology, USA), or monoclonal mouse anti-Ki-67 (1:50, clone MIB-1; Dako, Denmark) antibodies were applied on the slides.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Poorly differentiated synovial sarcoma is associated with high expression of enhancer of zeste homologue 2 (EZH2)

doi: 10.1186/1479-5876-10-216

Figure Lengend Snippet: Kaplan-Meier survival curves for high vs. low EZH2, HeK27me3, and Ki-67 score. Hazard ratios (HR) of the high-score groups with 95% confidence intervals are shown at the bottom.

Article Snippet: Monoclonal mouse anti-EZH2 (1:25, clone 11, BD Biosciences, USA), monoclonal rabbit anti-trimethyl-Histone H3 Lys27 (1:200, clone C36B11, Cell Signaling Technology, USA), or monoclonal mouse anti-Ki-67 (1:50, clone MIB-1; Dako, Denmark) antibodies were applied on the slides.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Poorly differentiated synovial sarcoma is associated with high expression of enhancer of zeste homologue 2 (EZH2)

doi: 10.1186/1479-5876-10-216

Figure Lengend Snippet: Association of high EZH2, H3K27me3, and Ki-67 scores with the risk of large tumor size and distant metastasis (NS: non-significant)

Article Snippet: Monoclonal mouse anti-EZH2 (1:25, clone 11, BD Biosciences, USA), monoclonal rabbit anti-trimethyl-Histone H3 Lys27 (1:200, clone C36B11, Cell Signaling Technology, USA), or monoclonal mouse anti-Ki-67 (1:50, clone MIB-1; Dako, Denmark) antibodies were applied on the slides.

Techniques:

Journal: BMC Cancer

Article Title: Enhancer of zeste homolog 2 (EZH2) expression is an independent prognostic factor in renal cell carcinoma

doi: 10.1186/1471-2407-10-524

Figure Lengend Snippet: EZH2 protein expression in renal cell carcinoma (RCC) and corresponding normal tissue . A Negative control: RCC tissue specimen (clear cell carcinoma), the anti-EZH2 antibody was omitted. B Normal adult kidney tissue, adjacent to the RCC tumor tissue shown in C. C Overview (left panel) and higher resolution (right panel) of the boxed area of an RCC sample (clear cell carcinoma), showing very high (> 50%) nuclear EZH2 expression (arrows). D Overview (left panel) and higher resolution (right panel) of an RCC sample (clear cell carcinoma) showing >5-25% nuclear EZH2 expression (arrows). E Overview (left panel) and higher resolution of an RCC sample (clear cell carcinoma) exhibiting 0% nuclear EZH2 staining. Scale bars, 5 μm.

Article Snippet: The TMA slides were dewaxed and rehydrated using xylene and a series of graded alcohols, followed by heat induced antigen retrieval using a target retrieval solution (S2031, DakoCytomation, Glostrup, Denmark) in a pressure cooker for 10 min. Immunohistochemical staining was performed on an automated staining system (Techmate 500, DakoCytomation) with a mouse monoclonal anti-EZH2 antibody (1:25, BD Transduction Laboratories, Franklin Lakes, NJ, USA) for 45 min [ ].

Techniques: Expressing, Negative Control, Staining

Journal: BMC Cancer

Article Title: Enhancer of zeste homolog 2 (EZH2) expression is an independent prognostic factor in renal cell carcinoma

doi: 10.1186/1471-2407-10-524

Figure Lengend Snippet: Correlation of nuclear EZH2 expression with clinicopathological characteristics

Article Snippet: The TMA slides were dewaxed and rehydrated using xylene and a series of graded alcohols, followed by heat induced antigen retrieval using a target retrieval solution (S2031, DakoCytomation, Glostrup, Denmark) in a pressure cooker for 10 min. Immunohistochemical staining was performed on an automated staining system (Techmate 500, DakoCytomation) with a mouse monoclonal anti-EZH2 antibody (1:25, BD Transduction Laboratories, Franklin Lakes, NJ, USA) for 45 min [ ].

Techniques: Expressing

Journal: BMC Cancer

Article Title: Enhancer of zeste homolog 2 (EZH2) expression is an independent prognostic factor in renal cell carcinoma

doi: 10.1186/1471-2407-10-524

Figure Lengend Snippet: Analysis of cancer specific survival (CSS) in RCC, patients without metastases

Article Snippet: The TMA slides were dewaxed and rehydrated using xylene and a series of graded alcohols, followed by heat induced antigen retrieval using a target retrieval solution (S2031, DakoCytomation, Glostrup, Denmark) in a pressure cooker for 10 min. Immunohistochemical staining was performed on an automated staining system (Techmate 500, DakoCytomation) with a mouse monoclonal anti-EZH2 antibody (1:25, BD Transduction Laboratories, Franklin Lakes, NJ, USA) for 45 min [ ].

Techniques:

Journal: BMC Cancer

Article Title: Enhancer of zeste homolog 2 (EZH2) expression is an independent prognostic factor in renal cell carcinoma

doi: 10.1186/1471-2407-10-524

Figure Lengend Snippet: Analysis of cancer specific survival (CSS) in RCC, patients with metastases

Article Snippet: The TMA slides were dewaxed and rehydrated using xylene and a series of graded alcohols, followed by heat induced antigen retrieval using a target retrieval solution (S2031, DakoCytomation, Glostrup, Denmark) in a pressure cooker for 10 min. Immunohistochemical staining was performed on an automated staining system (Techmate 500, DakoCytomation) with a mouse monoclonal anti-EZH2 antibody (1:25, BD Transduction Laboratories, Franklin Lakes, NJ, USA) for 45 min [ ].

Techniques: