Western Blot Analysis of Molecular Markers

Whole-cell extracts were obtained for SDS-PAGE electrophoresis as previously described (25 (link)). Primary antibodies used were rabbit anti-SMARCA4 (Abcam, ab110641), H3K27Me3 (Millipore, 07–449), Myc (Abcam, ab32072), mouse anti-EZH2 (BD Bioscience, 612667), vinculin (Sigma, v9264), Actin (Sigma, A5441), Histone H3 (Abcam, ab1791), MAP2 (Millipore, MAB3418), TUBB3 (Santa Cruz, sc-51670), BAD (Santa Cruz, sc-8044), p16 (Ventana, 725–4713) and CDKN1A/p21 (BD Pharmacogen, 556430) and horseradish peroxidase-conjugated anti-mouse and anti-rabbit IgG (Amersham). Chemiluminescence was used for detection (Perkin–Elmer Life Sciences). Actin or vinculin were used as protein loading controls.

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Wang Y., Chen S.Y., Colborne S., Lambert G., Shin C.Y., Santos N.D., Orlando K.A., Lang J.D., Hendricks W.P., Bally M.B., Karnezis A.N., Hass R., Underhill T.M., Morin G.B., Trent J.M., Weissman B.E, & Huntsman D.G. (2018). Histone deacetylase inhibitors synergizes with catalytic inhibitors of EZH2 to exhibit anti-tumor activity in small cell carcinoma of the ovary, hypercalcemic type. Molecular cancer therapeutics, 17(12), 2767-2779.

Publication 2018

H3K27Me3 mouse anti-EZH2 vinculin Histone H3 TUBB3 horseradish peroxidase-conjugated anti-mouse and anti-rabbit IgG

Actin Anti igg Antibodies Cell extracts Chemiluminescence Electrophoresis Ezh2 Histone h3 Horseradish peroxidase Map2 Mouse Protein Rabbit Sds page Smarca4 Tubb3 Vinculin

Corresponding Organization : University of British Columbia

Other organizations : Canada's Michael Smith Genome Sciences Centre, BC Cancer Agency, University of North Carolina at Chapel Hill, Translational Genomics Research Institute, Medizinische Hochschule Hannover

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Variable analysis

independent variables

SMARCA4 antibody
H3K27Me3 antibody
Myc antibody
EZH2 antibody
Vinculin antibody
Actin antibody
Histone H3 antibody
MAP2 antibody
TUBB3 antibody
BAD antibody
P16 antibody
CDKN1A/p21 antibody
Anti-mouse IgG
Anti-rabbit IgG

dependent variables

Protein expression levels

control variables

Actin
Vinculin

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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