Figure 8. Overexpression of STAT1 in OV3R-PTX-B4 cells. Cells were transfected with STAT1 plasmid (STAT1-Pls) or negative control vector (NC). The overexpression of STAT1 was confirmed by Western blot. (A) The expression of STAT1, CD44, CD133, NANOG, and OCT4 in STAT1-overexpressing cells. Representative images of blotting are shown. β-tubulin was used as a loading control. (B) Semi-quantitative analysis of the relative optical density of protein bands in A. (C) Spheroid formation assay of the cells. Cells were cultured in serum-free DMEM/F12 medium with EGF, bFGF, heparin, and B27 supplements for 11 days. Photos were taken every two days. (D) Measurement of the diameter of each spheroid in C. (E) Cell viability measurement by the CCK-8 assay. (F) Colony formation. A representative picture of the colonies is shown. (G) Number of colonies is shown. n = 3 independent experiments; *, P < 0.05; **, P < 0.01.