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. 2018 Jun 12;9(45):27835–27850. doi: 10.18632/oncotarget.25600

Figure 4. Apoptosis and survival signaling after L-OHP and CPT-11.

Figure 4

(A) Western blot analysis using antibodies against p53 and pro-apoptotic BAX and PIG-3 after treatment with 5 μM L-OHP or 10 μM CPT-11. (B) Immunodetection of NF-κB p65, RELB and anti-apoptotic survivin, XIAP, BCL-XL and MCL1; vinculin serves as loading control. (C) Effects of increasing doses L-OHP and CPT-11 on caspase-3 and PARP1 cleavage after 24 hours treatment; α-tubulin serves as loading control. (D) Cells were treated with a combination of L-OHP and the caspase-inhibitor Z-VAD-FMK (50 μM). Immunodetection of survivin, p53 and full-length caspase-3 was conducted. Detection of apoptosis was determined by cleavage products of caspase-3 and PARP1; β-actin serves as loading control. Please note: Figure 4A and 4B, as well as Supplementary Figure 2A show signals acquired by different detection methods, but originate from the same Western blots. This is due to a switch in the immunoblot chemiluminescence detection system from X-ray films (darker background) to a CCD camera system (Fusion Solo S, Vilber Lourmat; lighter background).