In (A), Proximity ligation assay. Cells were treated with X1 (10 μM) for 30 min and 60 min. Red dots indicate proximity of p-JNK and TOM20 (distance between the two proteins < 40 nm). Nuclei stained with DAPI (blue). In (B) The number of dots/cell were quantitated counting a minimum of 200 cells/experimental condition. Error bars represent ±SEM from three independent experiments, *p<0.05 vs vehicle. Scale bar, 20 μm. In (C and D), Huh7 cells incubated in HBSS were loaded with TMRM, as described in MATERIAL &METHODS. (C), cells were pretreated with Tat-sab KIM1 peptide (30 μM) for 1 h before treatment with X1 (10 μM) (upper panels) or sorafenib (5 μM) (lower panels). Image intensity was pseudocolored according to the reference bar. In (D), TMRM fluorescence was quantified after the various treatments compared to vehicle. *p<0.05 vs vehicle, #p<0.05 vs X1, §p<0.05 vs sorafenib from 3 independent experiments.