mouse monoclonal antibodies against ezh2  (Cell Signaling Technology Inc)

Journal: Breast Cancer Research : BCR

Article Title: BRCA1-deficient mammary tumor cells are dependent on EZH2 expression and sensitive to Polycomb Repressive Complex 2-inhibitor 3-deazaneplanocin A

doi: 10.1186/bcr2354

Figure Lengend Snippet: Ezh2 expression is elevated in BRCA1-deficient primary mouse mammary tumors. (a) mRNA levels of Ezh2 in BRCA1-deficient ( K 14cre; B rca 1 F / F ; p 53 F / F (KB1P)) and BRCA1-proficient ( K 14cre;Brca1 w . t / w . t ; p 53 F / F (KP)) mammary tumors analyzed by microarray analysis. The mean (± standard error of the mean) log2 ratio of Ezh2 expression in 21 KP tumors is -0.036 (± 0.067) and 0.497 (± 0.054) in 32 KB1P tumors. The Ezh2 expression is significantly higher in KB1P tumors compared with KP tumors (*Wilcoxon exact test). (b) EZH2 protein levels in two independent primary KB1P and in two independent primary KP tumors detected by immunohistochemistry (scale bar represents 100 μm), representative of a total of four tumors analysed for each genotype. (c) Quantification of EZH2 immunohistochemistry shown in b (* Wilcoxon exact test).

Article Snippet: The samples were incubated overnight with a mouse monoclonal antibody against EZH2 (1:100, BD Biosciences San Jose, California, USA).

Techniques: Expressing, Microarray, Immunohistochemistry

Journal: Breast Cancer Research : BCR

Article Title: BRCA1-deficient mammary tumor cells are dependent on EZH2 expression and sensitive to Polycomb Repressive Complex 2-inhibitor 3-deazaneplanocin A

doi: 10.1186/bcr2354

Figure Lengend Snippet: EZH2 is overexpressed in BRCA1-deficient human breast tumors. (a) The mean (± standard error of the mean) log10 ratio of EZH2 expression in human breast cancer samples is -0.15 (± 0.041; >five-year survival, good prognosis), 0.028 (± 0.046;

Article Snippet: The samples were incubated overnight with a mouse monoclonal antibody against EZH2 (1:100, BD Biosciences San Jose, California, USA).

Techniques: Expressing, Immunohistochemistry

Journal: Breast Cancer Research : BCR

Article Title: BRCA1-deficient mammary tumor cells are dependent on EZH2 expression and sensitive to Polycomb Repressive Complex 2-inhibitor 3-deazaneplanocin A

doi: 10.1186/bcr2354

Figure Lengend Snippet: EZH2 is required for survival of BRCA1-deficient but not BRCA1-proficient tumors cells. (a) Ezh2 mRNA expression measured by quantitative RT-PCR in BRCA1-deficient cells (in red, K 14cre; B rca 1 F / F ; p 53 F / F (KB1P)) and in BRCA1-proficient cells (in blue, K 14cre;Brca1 w . t / w . t ; p 53 F / F (KP)) after treatment with siRNAs against Ezh2 or 5 μM 3-deazaneplanocin A (DZNep; shown as fold induction relative to Hprt ). (b) EZH2 levels analysed by western blot after KB1P and KP cells were treated with siRNAs targeting Ezh2 or non-targeting siRNAs (siNTC) and 5 μM DZNep or vehicle (dimethyl sulfoxide (DMSO)) for the indicated period. (c) Phase-contrast images of BRCA1-deficient and BRCA1-proficient cells treated for 48 hours with control (ctrl) siRNAs, siRNAs against Ezh2 or 5 μM DZNep (original magnification 10×). (d) Growth curves of KB1P (depicted in red) and KP cell lines (depicted in blue) treated with control (ctrl) siRNAs, siRNAs against Ezh2 or 5 μM DZNep. Data measured by Cell Titer Blue and represented as the mean ± standard error of the mean (three independent experiments). The depicted cell lines are representative for all three KB1P and KP cell lines.

Article Snippet: The samples were incubated overnight with a mouse monoclonal antibody against EZH2 (1:100, BD Biosciences San Jose, California, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: BRCA1-deficient mammary tumor cells are dependent on EZH2 expression and sensitive to Polycomb Repressive Complex 2-inhibitor 3-deazaneplanocin A

doi: 10.1186/bcr2354

Figure Lengend Snippet: Chemical EZH2-inhibitor DZNep selectively kills BRCA1-deficient tumor cells. (a) Representative growth inhibition curves for BRCA1-deficient cell lines ( K 14cre; B rca 1 F / F ; p 53 F / F (KB1P), in blue) and BRCA1-proficient cell lines ( K 14cre;Brca1 w . t / w . t ; p 53 F / F (KP), in red) treated with 3-deazaneplanocin A (DZNep). A serial dilution of DZNep was added to the cells and cell viability was measured five days later (each data point represents the mean ± standard error of the mean (SEM) of three independent experiments). (b) Representative growth inhibition curves for BRCA1-deficient cell lines (KB1P, in blue) and BRCA1-proficient cell lines (KP, in red) treated with trichostatin A (TSA). A serial dilution of TSA was added to the cells and cell viability was measured five days later (mean ± SEM).

Article Snippet: The samples were incubated overnight with a mouse monoclonal antibody against EZH2 (1:100, BD Biosciences San Jose, California, USA).

Techniques: Inhibition, Serial Dilution

Journal: Breast Cancer Research : BCR

Article Title: BRCA1-deficient mammary tumor cells are dependent on EZH2 expression and sensitive to Polycomb Repressive Complex 2-inhibitor 3-deazaneplanocin A

doi: 10.1186/bcr2354

Figure Lengend Snippet: Restoration of BRCA1 partially rescues from sensitivity to DZNep. (a) Detection of the human BRCA1 allele by PCR amplification of exon 11 in the reconstituted subclones KB1PR-3.12 E3 and F4. The human breast cancer cell line T47D was used as a positive control. (b) EZH2 protein levels of (untreated) K K 14cre; B rca 1 F / F ; p 53 F / F (KB1P), KB1PR and K 14cre;Brca1 w . t / w . t ; p 53 F / F (KP) cell lines were analysed by western blotting. Corresponding Ezh2 mRNA levels of KB1P, KB1PR and KP cell lines were measured by quantitative RT-PCR (shown as fold induction relative to Hprt ). (c) Representative growth inhibition curves for BRCA1-deficient cell lines (KB1P, in blue), BRCA1-proficient cell lines (KP, in red) and two clones of a BRCA1 -reconstituted cell line (KB1PR E3 and F4, in purple) treated with 3-deazaneplanocin A (DZNep). A serial dilution of DZNep was added to the cells and cell viability was measured five days later (mean ± standard error of the mean). (d) EZH2 protein levels of hBRCA1 -reconstituted KB1P cells 48 hours after treatment with DZNep or siRNAs targeting EZH2.

Article Snippet: The samples were incubated overnight with a mouse monoclonal antibody against EZH2 (1:100, BD Biosciences San Jose, California, USA).

Techniques: Amplification, Positive Control, Western Blot, Quantitative RT-PCR, Inhibition, Clone Assay, Serial Dilution

Journal: Stem Cells and Development

Article Title: Identification of Novel EZH2 Targets Regulating Osteogenic Differentiation in Mesenchymal Stem Cells

doi: 10.1089/scd.2015.0384

Figure Lengend Snippet: Table 3.

Article Snippet: Antibodies against anti-rabbit H3K27me3 (1 μg, Merck, Millipore 07-449, Bayswater, VIC, AUS, http://www.merck.com.au/ ), anti-human EZH2 mouse monoclonal (1 μg, Millipore AC22 17-662), anti-human H3K4me3 rabbit polyclonal (1 μg, Abcam ab8580-100), and anti-human IgG rabbit polyclonal control (1 μg Millipore, Bayswater, VIC, AUS) were used for immunoprecipitation.

Techniques: