Fig. 5. The effect of SOX6-induced autophagy in the sensitivity of cervical cancer cells to cisplatin treatment in vitro and in vivo.

A Top 20 Gene Ontology (GO)-enriched terms of MAP4K4 gene. *indicated the terms related to apoptosis and cell death. B Flow cytometry analysis on the apoptosis of HeLa-HA-SOX6-tet and HeLa-HA-SOX6ΔHMG-tet cells with or without cisplatin (20 μM) treatment for 48 h. C The percentage of apoptotic cells in flow cytometry analysis. D Western blotting analysis on the protein levels of PARP and caspase 9 in HeLa-HA-SOX6-tet cells with or without cisplatin (20 μM) treatment. α-tubulin protein was used as the internal control. E Experimental design of the steps used to produce xenograft in BALB/c nude mice. F Subcutaneously injection of the HeLa-HA-SOX6-tet cells into the left flank of nude mice, which were subsequently divided into two groups at 1-week post-injection and were intraperitoneally injected with Dox (20 mg/kg, PBS as solvent control) every day and cisplatin (3 mg/kg, saline as solvent control) every other day for the next 2 weeks. The relative sizes of the tumors were represented by the ratio of the tumor size in cisplatin group to those in saline group. G Immunofluorescence staining of SOX6 (green) and p62/SQSTM1 protein (red) in the frozen section of xenograft tumor tissues. H Relative expression level of p62/SQSTM1 indicated by fluorescent density. IOD integral optical density. I Immunofluorescence staining of TUNEL (green) in the frozen section of xenograft tumor tissues. J The percentage of TUNEL-positive cells in the frozen section of xenograft tumor tissues. Data are mean ± SEM of three independent experiments or at least three fields scored. (*P < 0.05, **P < 0.01, ***P < 0.001, NS non-significant, Student’s t-test, two tails for F, one-way ANOVA and post hoc Tukey tests for C, H, J).