Immunoblotting of Cancer Signaling Proteins

Immunoblotting was performed as described previously (48 (link)). The following primary antibodies were used: anti-BRD4 antibody (A301-985A100, Bethyl Laboratories), anti-RET antibody (ab134100, Abcam), anti-RET (phospho Y1015) antibody (ab74154, Abcam), anti-Raf1 antibody (A19638, Abclonal), anti-phospho-Raf1-S259 antibody (AP1012, Abclonal), anti-MEK1/MEK2 antibody (A4868, Abclonal), anti-phospho-MAP2K1-S217/MAP2K2-S221 antibody (AP0209, Abclonal), anti-ERK1/2 antibody (A4782, Abclonal), anti-phospho-ERK1-T202/Y204 + ERK2-T185/Y187 antibody (AP0472, Abclonal), anti-p90Rsk antibody (A4695, Abclonal), anti-phospho-P90RSK-S380 antibody (AP0562, Abclonal), anti-ERα antibody (ab32063, Abcam), anti-phospho-ERα (Ser167) antibody (64508s, Cell Signaling Technology), anti-phospho-ERα (Ser118) antibody (ab32396, Abcam), anti-CCND1 antibody (ab40754, Abcam), anti-c-MYC antibody (SC-40, Santa Cruz Biotech) and anti-ACTIN antibody (8432, Cell Signaling Technology).

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Zheng Z.Z., Xia L., Hu G.S., Liu J.Y., Hu Y.H., Chen Y.J., Peng J.Y., Zhang W.J, & Liu W. (2022). Super-enhancer-controlled positive feedback loop BRD4/ERα–RET–ERα promotes ERα-positive breast cancer. Nucleic Acids Research, 50(18), 10230-10248.

Publication 2022

ab134100 A4868 AP0209 A4782 AP0472 ab32063 ab32396 ab40754 SC-40

A301 Actin Anti antibody Anti c antibody Antibodies Antibody Brd4 Ccnd1 Erk1 Erk2 Map2k1 Map2k2 Mek2 P90rsk Phospho Raf1

Corresponding Organization : First Affiliated Hospital of Gannan Medical University

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Variable analysis

independent variables

Immunoblotting protocol

dependent variables

BRD4 protein expression
RET protein expression
Phosphorylated RET (Y1015) protein expression
Raf1 protein expression
Phosphorylated Raf1 (S259) protein expression
MEK1/MEK2 protein expression
Phosphorylated MEK1 (S217)/MEK2 (S221) protein expression
ERK1/2 protein expression
Phosphorylated ERK1 (T202/Y204) and ERK2 (T185/Y187) protein expression
P90Rsk protein expression
Phosphorylated P90RSK (S380) protein expression
ERα protein expression
Phosphorylated ERα (S167) protein expression
Phosphorylated ERα (S118) protein expression
CCND1 protein expression
C-MYC protein expression
ACTIN protein expression

control variables

Antibodies used for immunoblotting

Annotations

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