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. 2022 Sep 20;12:976292. doi: 10.3389/fonc.2022.976292

Figure 5.

Figure 5

Combination of KP372-1 with PARP inhibitor induces cell autophagy and cell apoptosis. Cells were pre-treated ± rucaparib (0.4 µM or 15 µM, 2 h), then exposed to KP372-1 ± rucaparib or KP372-1 ± rucaparib ± Bafilomycin A1 (Baf A1) for 2 h followed by washing and replacing media; cells treated with Baf A1 (0.05 µM) were kept with Baf A1 for 24 h; positive control cells were exposed to Staurosporine (STS, 1 μM) for 18 h; then cells including debris in media were finally collected at indicated timepoints and examined for: (A) Annexin-V/7-AAD staining to determine cell death way via flow cytometry, early apoptosis part indicated by Annexin-V+/7-AAD- was quantified on the left pannel; (B) PARP1 and cleaved caspase 7 alterations in A549 cells; (C) Levels of cleaved caspase 7 in MCF-7 cells; (D, E) LC3 I/II and p62 levels in A549 cells. Results were separately repeated at least three times and protein levels were quantified. (A) Error bars are means ± SD. ** P < 0.01 *** P < 0.001, comparing each group with control (DMSO) treatment (t tests).