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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: ROS-p53-cyclophilin-D signaling mediates salinomycin-induced glioma cell necrosis

Fig. 4

Salinomycin induces Cyp-D and p53 interaction in mitochondria. Expression of p53, Cyp-D and tubulin expression in stable U87MG cells with scramble-shRNA or p53-shRNA was shown (a). Above cell lines were treated with salinomycin (Sali, 5 μM) for 72 h, cell necrosis and apoptosis were tested by FACS (b). Cultured U87MG glioma cells were treated with indicated concentration of salinomycin (Sali) for 6 h, expression of p53, p-p53 (Ser 15) and Cyp-D in the mitochondrial fraction was tested by Western blots (c). Scramble- or p53- sh-RNA infected U87MG cells, pre-incubated with CsA (0.1 μM, 1 h), were treated with salinomycin (Sali, 5 μM) for 6 h, mitochondrial association of p53 and Cyp-D was tested (d), expression of above protein in whole cell lysates (WCL) was shown in input (d), mitochondrial membrane potential was also tested by JC-10 fluorescence assay (e). U87MG glioma cells were treated with salinomycin (Sali, 5/10 μM), or TMZ (5 mM) for 6 h, p-p53 and regular p53 were tested by Western blots (f). Error bars indicate SD. Experiments were repeated three times in this figure, and similar results were obtained. *p < 0.05 (b and e)

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