a, HeLa cells were treated for 12 h with 10 μM 3 in the absence and presence of 40 μM ZVAD-FMK. The indicated proteins were immunoblotted using the corresponding antibodies. ‘Un’ indicates no treatment of cells with 3. b, HeLa cells stably expressing mRFP-EGFP-LC3 were treated for 24 h with 10 μM 3 in the absence and presence of 40 μM ZVAD-FMK. Cell images of EGFP and mRFP, obtained using confocal fluorescence microscopy, were merged (scale bar, 10 μm). ‘Un’ indicates no treatment of cells with 3. c, Flow cytometry of HeLa cells treated with 10 μM 3 or/and 5 nM BfA1 for 12 h and then stained with JC-1 (left) or fluorescein-annexin V (right). ‘Untreated’ indicates no treatment of cells with 3 and BfA1. d, Caspase activities of lysates of HeLa cells treated with 10 μM 3 or/and 5 nM BfA1 for 12 h were measured using 200 μM acetyl-DEVD-pNA in the absence (grey bars) and presence (black bars) of 20 μM Ac-DEVD-CHO (mean ± s.d., n = 3). ‘Un’ indicates no treatment of cells with 3 and BfA1. e, HeLa cells were treated with 10 μM 3 or/and 5 nM BfA1 for 12 h. The indicated proteins were then immunoblotted by using the corresponding antibodies. ‘Un’ indicates no treatment of cells with 3 and BfA1.