Fig. 2

CICD does not trigger proliferation of melanoma cancer cells. a Working model for testing the effect of apoptotic and CICD conditioned media on the proliferation of neighboring cells. b-c Incucyte analysis for the proliferation of WM115 H2B-mCherry cells grown in the presence of APO (b) or CICD (c) conditioned media, obtained 24 h after triggering either apoptosis or CICD. n = 4–5 independent experiments; mean values +/− s.e.m. d Same as in (b-c), while this time cell proliferation was assessed by quantifying the optical density (O.D.) of methylene blue staining of cells grown in either apoptotic or CICD media. e Representative images of methylene blue staining. f Compensatory proliferation tested and quantified by clonogenic survival assay performed using the same conditions described in (b). g Parental WM115 cells were incubated with doxycycline (1 μg/ml) and the conditioned media was then added on H2B-mCherry expressing WM115 cells while clonogenic survival was assessed. Actinomycin D (ActD) treatment was used as negative control. h Incucyte analysis for the cell death induction (SYTOX Green exclusion) triggered by CICD conditioned media in WM115 H2B-mCherry cells. Actinomycin D treatment (1 μM) is used as positive control for cell death induction. A representative experiment is shown. i Immunoblotting for ZEB-1-MITF transcription factors axis of WM115 cells grown in APO and CICD conditioned media for 48 h. Actin was used as loading control