Fig. 1

SGK3 is preferentially activated in liver CSCs. a MHCC-97H cells were cultured in monolayer or ultra-low attachment conditions. The mRNA expression of liver CSC-related genes and SGK3 in spheroids and attached cells was compared by RT-PCR. b Western blot analysis for levels of CD133, active Akt (phosphorylated at Ser473)/total Akt and active SGK3 (phosphorylated at Thr320)/total SGK3 between spheroids and attached cells. c Flow cytometry analysis of CD133+ cell distribution in CD133- and CD133+ cells isolated using CD133 MicroBead Kit. d Representative images of CD133+ and CD133- cells sorted from MHCC97H HCC cells cultured in serum-free culture medium after 7 days. Scale bars, 100 μm. e CD133+/CD133- cells were subcutaneously injected into 6-week-old female athymic nude mice, and tumourigenicity was evaluated 5 weeks post-inoculation. f Levels of CD133, active Akt (phosphorylated at Ser473)/total Akt and active SGK3 (phosphorylated at Thr320)/total SGK3 were compared by western blot analysis between CD133+ and CD133- cells. β-actin was used as a loading control. All experiments were performed in triplicate. *P < 0.05