Fig. 1

Efficient engagement of either apoptosis or CICD in melanoma cells. a. In vitro system of triggering either apoptosis or CICD in melanoma cells based on doxycycline-mediated rapid expression of the pro-apoptotic BAX protein. b Apoptosis (1 μg/ml DOX-treated cells) or CICD (DOX and Q-VD-OPh treated cells) was engaged in WM115 for 24 h followed by immunoblotting for BAX, caspase-3, PARP1 and actin as loading control. c WM115 cells were treated as in (b) and cell viability was measured by SYTOX Green exclusion in an Incucyte Imager. A representative experiment is shown. d Efficacy of CRISPR/Cas9-mediated APAF-1 KO in WM115 cells. e Validation of apoptosis and CICD induction in APAF-1 KO WM115 cells. DOX treatment and immunoblotting was done as described in (b). f Representative SYTOX Green positive staining for either apoptotic or cells undergoing CICD at 24 h. g Cell death kinetics for apoptosis and CICD triggered in the context of APAF-1 KO. A representative experiment is shown. h-i. Immunofluorescence representative images (h) for the release of cytochrome c and quantification (i)