Fig. 4
Effects of CPP on cell viability, DNA damage response, cell death and antioxidant system. (a) HuCCT-1 and EGI-1 CCA cells viability is quantified 24 h after CPP exposure for 0.5, 1, 2, 3 and 5 min by crystal violet assay. (b-c) CCA cells are treated with Pz-DBD for 2 and 3 min for HuCCT-1 – 3 and 5 min for EGI-1 and analyses are performed 24 h post-treatment. (b) Representative images of γH2AX analyzed by immunofluorescence in cells. (c) Representative images of western blot analysis of γH2AX, phosphorylated and total Chk1, phosphorylated and total p53, cleaved caspase-3 and cleaved PARP. The samples derive from the same experiment and gels/blots are processed in parallel. Blots are cropped; original uncropped blots are presented in Supplementary materials. (d) Expression of MSRB3, SOD1, SOD2, CAT2 and HMOX1 at mRNA level by RT-QPCR. Values are expressed as means ± s.e.m. from at least three independent cultures. Significant differences for *, p < 0.05; **, p < 0.01; ***, p < 0.001 are compared with untreated cells (0 min).
