Fig. 2

Hes1 ectopic expression promotes CSC characteristics and resistance to cisplatin. a Schematic illustration describing the transduction procedures of OV2774 and SKOV3 cells with the pHes1dEGFP reporter and data analysis. b Transduction of cells with the pHes1d2EGFP reporter and representative photomicrographs of parental cells and spheres. c Parental and Hes1GFP(+) cells were cultured, dissociated from spheres, labelled with anti-Hes1 antibody and isotype control and analyzed by flow cytometry. d Cell lysates were prepared from the indicated cells and analyzed by immunoblotting. e Total mRNA was purified and the level of the HES1 mRNA was assessed by qRT-PCR, (n = 3, mean +/− SD; Students t-Test; ** p 0.01). f Cell growth of the indicated cells was analyzed by the WST1 assay. (n = 3, mean +/− SD; Students t-Test; ** p 0.01). g Sphere formation ability of Hes1GFP(+) and GFP(−) cells was analyzed at serial generations. Photographs are from secondary cultures. Sphere numbers were counted and presented as bar graph (n = 3, mean +/− SD; Students t-Test; ** p 0.01). h Cells were treated with increasing concentrations of cisplatin and cytotoxicity was assessed by the WST1 assay. (n = 3, mean +/− SD; Students t-Test; * p 0.05). i Total RNA was purified from Hes1-GFP-positive and GFP-negative cells, and then the expression of the indicated genes was assessed by qRT-PCR (n = 3, mean +/− SD; Students t-Test;*p 0.05, ** p 0.01). j Transfected cells were FACS-sorted as described in (a; b). Sorted GFP(+/−) cells were transplanted subcutaneously in mice (n = 5/group). Resected tumors were photographed 8 weeks after implantation. Bar graph showing the tumor weight in each group (Students t-Test; mean +/− SD **p 0.01)