Cells were lysed with chilled RIPA buffer (Sigma-Aldrich, MO, USA). Samples were centrifuged (12,000×g), and lysates were added to LDS sample buffer. Samples were separated via SDS-PAGE and transferred to PVDF membranes (Bio-Rad, CA, USA) that were then blocked using 5% nonfat milk, incubated overnight with primary antibodies at 4 °C overnight, and incubated with a secondary antibody (1:1000; CST, MA, USA). Protein was then detected with a chemiluminescent detection system (Bio-Rad).
Primary antibodies used in this study were specific for the following: NRG1 (ab 191139, Abcam, MA; 1:500), ErbB2 (ab 241325, Abcam; 1:1000), p-Akt (ab 241325, Abcam; 1:1000), p-ERK (ab229912, Abcam; 1:1000), p-mTOR (ab 109268, Abcam; 1:500), GAPDH (ab181603, Abcam; 1:2000), p-P65(ab 53489, Abcam; 1:1000), P65 (ab 32536, Abcam; 1:5000), LaminB (ab 16048, Abcam; 1:1000).
Liu Y., Zhang N., Wang Y., Zuo J., Wang J., Chu Y, & Ye Y. (2023). Chorionic villus-derived mesenchymal stem cell-mediated NRG1 upregulation promotes HTR-8/SVneo cells proliferation through the activation of the NF-κB signaling pathway. Heliyon, 9(8), e18245.
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