Immunoblot Analysis of Cell Lysates

Total lysates prepared from subconfluent cells as described^{24 (link)} were subjected to immunoblot analysis using anti-AR (H-280, 1:100 dilution; Santa Cruz Biotechnology), anti-PSA (5365, 1:1000 dilution; Cell Signaling Technology, Danvers, MA, USA), anti-NKX3.1 (83700, 1:1000 dilution; Cell Signaling Technology), anti-β-actin (1:100,000 dilution; Sigma), anti-MUC1-C (HM-1630-P1ABX, 1:400 dilution; Thermo Fisher Scientific, Waltham, MA, USA), anti-EZH2 (5246, 1:1000 dilution; Cell Signaling Technology), anti-MYCN (9405, 1:1000 dilution; Cell Signaling Technology), anti-BRN2 (12137, 1:1000 dilution; Cell Signaling Technology), anti-MYC (ab32072, 1:1000 dilution; Abcam, Cambridge, MA), anti-SOX2 (3579, 1:1000 dilution; Cell Signaling Technology), anti-ASCL1 (GTX129189, 1:1000 dilution; GeneTex, Irvine, CA, USA), anti-AUROKA (ab1287, 1:4000 dilution; Abcam), anti-SYP (MA5-16402, 1:200 dilution; Thermo Fisher Scientific), anti-KLF4 (12173, 1:1000 dilution; Cell Signaling Technology) and anti-OCT4 (2750, 1:1000 dilution; Cell Signaling Technology).

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Yasumizu Y., Rajabi H., Jin C., Hata T., Pitroda S., Long M.D., Hagiwara M., Li W., Hu Q., Liu S., Yamashita N., Fushimi A., Kui L., Samur M., Yamamoto M., Zhang Y., Zhang N., Hong D., Maeda T., Kosaka T., Wong K.K., Oya M, & Kufe D. (2020). MUC1-C regulates lineage plasticity driving progression to neuroendocrine prostate cancer. Nature Communications, 11, 338.

Publication 2020

anti-AR (H-280 anti-PSA anti-NKX3.1 anti-β-actin anti-MUC1-C (HM-1630-P1ABX anti-EZH2 anti-BRN2 anti-SOX2 ab1287 anti-KLF4 anti-OCT4

Actin Brn2 Cells Dilution Ezh2 Immunoblot analysis Klf4 Muc1 Mycn Oct4 Sox2

Corresponding Organization :

Other organizations : Dana-Farber Cancer Institute, Harvard University, University of Chicago, Roswell Park Comprehensive Cancer Center, Keio University, NYU Langone Health

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Variable analysis

independent variables

Independent variables not explicitly mentioned.

dependent variables

AR protein levels
PSA protein levels
NKX3.1 protein levels
MUC1-C protein levels
EZH2 protein levels
MYCN protein levels
BRN2 protein levels
MYC protein levels
SOX2 protein levels
ASCL1 protein levels
AUROKA protein levels
SYP protein levels
KLF4 protein levels
OCT4 protein levels

control variables

Total lysates prepared from subconfluent cells as described in reference 24

positive controls

β-actin

Annotations

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