Figure 4.

miR-361-3p directly targeted SOX1 in liver T-ICs. A. Schematic diagram of luciferase plasmids of SOX1 3'-UTR wild-type (WT) and SXO1 3'-UTR mutant (MUT). B. The SOX1 mRNA expression in Huh7/HepG2 miR-361-3p sponge and control cells were detected by real-time PCR. C. The SOX1 mRNA expression in Huh7/HepG2 miR-361-3p mimic and control cells were detected by real-time PCR. D. The SOX1 protein expression in Huh7/HepG2 miR-361-3p sponge and control cells were detected by western blot. E. The SOX1 protein expression in Huh7/HepG2 miR-361-3p mimic and control cells were detected by western blot. F. HCC cells were stably transfected with SOX1 3'-UTR wild-type (WT) and SXO1 3'-UTR mutant (MUT) luciferase reporter for 24 hours. The luciferase activity was measured as described in “materials and methods”. G. A negative correlation was observed between miR-361-3p and SOX1 expression in 50 clinical samples of HCC (R² = 0.734, P < 0.05, by Spearman's correlation analysis). H. Huh7/HepG2 miR-361-3p sponge and control cells were transfected with siRNA SOX1 or its vector control. The stable transfectants were identified by Western Blotting. I. Huh7/HepG2 miR-361-3p sponge and control cells transfected with shRNA SOX1 and then subjected to spheroids formation assay. J. Huh7/HepG2 miR-361-3p sponge and control cells transfected with shRNA SOX1 and then subjected to in vitro limiting dilution assay.