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. 2020 May 4;11(5):308. doi: 10.1038/s41419-020-2502-4

Fig. 5. M50054 inhibits active Apaf-1.

Fig. 5

The effect of M50054 on caspase-9 (a) and -3 (b) activity induced by 1 mM dATP and 1.6 µM cytochrome c (dATP/Cc). After 15 min incubation with dATP/Cc the activity of caspase-9 like activity was measured with LEHD-pNA and the activity of caspase-3 activity measured with DEVD-AFC. c Immunoblotting to assess the effect of M50054 on dATP and cytochrome c induced processing of caspase-9 and -3. The results of (a), (b) and (c) are from one cell extract and are typical of three independent experiments. d The effect of M50054 (500 µM) on Apaf-1:Apaf-1 interactions were assessed using a split luciferase complementation assay. e The effect of M50054 (500 µM) on luciferase activity relative to DMSO (0.1% v/v). f The binding of M50054 to inactive and active Apaf-1 was assessed using a thermal stability assay. g The binding of M50054 to cytochrome c was assessed using a thermal stability assay. 10 mM ATP was used as a positive control. The data are the mean ± SD of three independent experiments (*P < 0.05).