Mitochondrial Cytochrome C and BAX Regulation

Jurkat cells at 2 ×10⁶ cells/mL were incubated in the presence of DMSO or 50 μM BIP-V5 inhibitor^{54 (link)} (Yuanye Biotechnology Co., Ltd, 579492-81-2) for 24 h. Two groups of above treated cells were each divided into four groups and incubated in the presence of DMSO, venetoclax at 2.5 μM, cp4-TAT at 20 μM, or cp5-TAT at 15 μM in serum-free RPMI medium for 4 h. Then the medium was supplemented with FBS to 10% and incubated for another 8 h. After being treated by venetoclax or CPs for totally 12 h, cell were processed with the Mitochondrial Separation Kit (Beyotime, C3601) to extract mitochondria. Cytochrome C in mitochondria and BAX proteins in the whole cells were detected by Western blot.