Figure 5.

LINC00337 regulates CSC-like properties of CD44⁺/CD24^low/−SFCs via miR-145. CD44⁺/CD24^low/−SFCs were transfected with miR-145 mimic, mimic control, miR-145 inhibitor, inhibitor control, sh-LINC00337 plus miR-145 inhibitor, and scramble shRNA plus inhibitor control for 48 h. (A) Statics of spheres per 1,000 cells. (B) Views and numbers of colonies in CD44⁺/CD24^low/−SFCs, examined by soft-agar colony formation assays. (C) CCK-8 assays were performed to evaluate the viability of CD44⁺/CD24^low/−SFCs. (D) IC50 values of cisplatin, doxorubicin and epirubicin in CD44⁺/CD24^low/−SFCs. (E) Cell cycle distribution of CD44⁺/CD24^low/−SFCs was determined by flow cytometric analysis of PI staining. (F) Cell apoptosis of CD44⁺/CD24^low/−SFCs was determined by flow cytometric analysis of Annexin V-FITC/PI double staining. (G) The mRNA expressions of MDR-1, Nanog, Sox2, and Oct4 in CD44⁺/CD24^low/−SFCs were determined by the qRT-PCR analysis, normalized to GAPDH expression. (H) Representative Western blots of MDR-1, Nanog, Sox2, and Oct4 and their quantitative analyses in CD44⁺/CD24^low/−SFCs, normalized to GAPDH expression. Data are shown as mean ± standard deviation of three technical replicates and analyzed by one-way ANOVA, for (C), by two-way ANOVA for (C). *p < 0.05 vs. mimic control. **p < 0.01 vs. mimic control. ^#p < 0.05 vs. inhibitor control. ^##p < 0.01 vs. inhibitor control.