Fig. 2.

STON2 knockdown promotes stem-like properties of ovarian cancer cells. a 3AO and Caov3 cells were transfected with a STON2-specific shRNA for 48 h, and cultured in spheroid culture conditions for 7 days. The expressions of STON2 and the CSC-related markers NANOG and c-MYC were detected by immunoblot analysis. b Quantitation and representative images of sphere-formation (sphere > 50 μm), Scale bars, 50 μm. c The effect of STON2 knockdown on the CD44⁺CD24⁻ phenotype was analyzed using FCM. d Immunoblot analysis of EMT-related markers (E-cadherin, N-cadherin, and fibronectin). e qPCR analysis of EMT-related markers (E-cadherin, N-cadherin, and fibronectin). f, g 3AO cells were transfected with shNC or shSTON2 for 48 h, cultured in spheroid culture conditions for 7 days, and then injected into NOD/SCID mice (n = 5 in each group). All mice were sacrificed at week 4 and the tumor incidence evaluated (left panel). Subcutaneous tumors are shown (right panel) (f).Tumor diameters of xenografts from mice injected with 1 × 10⁵ cells were measured at a regular intervals of 1 week for up to 4 weeks and the tumor volume was calculated (g). Data represents mean ± S.E. of three independent experiments. The level of significance is indicated by *P < 0.05, **P < 0.01, ***P < 0.001