Protein extraction and western blot (WB) analysis were performed as previously described [14 (link)]. Briefly, cells were washed with PBS and lysed with lysis buffer on ice for 20 min. The total cell protein concentration was detected using the BCA Protein Assay Kit. The total protein (20 μg) was separated using SDS-PAGE (Invitrogen) and transferred to a PVDF membrane (Roche). The membrane was blocked with 5% bovine serum albumin (0.1%) in TBS-Tween and incubated against the required antibody.
The primary antibodies Bax (5023, Cell Signaling Technology, USA), Bcl2 (ab196495, Abcam, USA), cleaved caspase-3 (29034, Signalway Antibody, USA), BTG2 (A9848, ABclonal), GAPDH (5174, Cell Signaling Technology), TSG101 (14497, Proteintech, United States), CD63 (25682, Proteintech), CD81 (66866, Proteintech), and horseradish peroxidase-conjugated secondary antibody (Santa Cruz) were used. Bands were visualized using enhanced chemiluminescence reagents and analyzed using a gel documentation system (Bio-Rad Gel Doc1000 and Multi-Analyst version 1.1).
The primary antibodies Bax (5023, Cell Signaling Technology, USA), Bcl2 (ab196495, Abcam, USA), cleaved caspase-3 (29034, Signalway Antibody, USA), BTG2 (A9848, ABclonal), GAPDH (5174, Cell Signaling Technology), TSG101 (14497, Proteintech, United States), CD63 (25682, Proteintech), CD81 (66866, Proteintech), and horseradish peroxidase-conjugated secondary antibody (Santa Cruz) were used. Bands were visualized using enhanced chemiluminescence reagents and analyzed using a gel documentation system (Bio-Rad Gel Doc1000 and Multi-Analyst version 1.1).
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