Fig. 4

Caspase inhibitors failed to block AIF translocation in HaCaT cells treated with H₂O₂. a Annexin V-PI combined with flow cytometry was used to detect apoptosis in HaCaT cells treated with DPQ and Z-VAD-fmk. b PI- and AnnexinV-positive cells had lower expression levels than the H₂O₂ group(^#P < 0.01) but not the Z-VAD-fmk treatment. c–f Western blot of AIF translocation by nuclear fractionation in HaCaT cells 4 h after H₂O₂ treatment with 30 μM DPQ or 100 μM Z-VAD-fmk. Mitochondrial AIF was upregulated in the DPQ group but downregulated in the Z-VAD-fmk group. Nuclear AIF and PARP-1 were downregulated in the DPQ group. g, h PAR in the whole protein was downregulated for the DPQ group but upregulated for the Z-VAD-fmk group. Relative to the control: ^**P < 0.01; compared to H₂O₂: ^##P < 0.01. Data are representative of three independent experiments (means ± SD). i, j Fluorescence microscopy of AIF and cleavedcaspase-3 for the various treatments. Scale bar = 200 μm