Figure 4

SOX9 silencing impairs self-renewal of SW620 cells.

(A) Representative immunoblots of SOX9 and BMI1 protein expression in empty vector (pLKO) and shSOX9 SW620 cells (n ≥ 3). (B) SOX9 mRNA levels in shSOX9 SW620 relative to pLKO cells (n = 3). (C) Plot of the number of primary and secondary tumorspheres in the indicated conditions (n ≥ 5). (D) Tumor initiation ability of the indicated cellular genotypes. Frequency of tumors formed in immunocompromised mice after subcutaneous injection of 1·10⁶ cells per injection. (E) Tumor volume calculated at the indicated time points in nude mice injected with pLKO or two independent shSOX9 constructs (shSOX9 and shSOX9-2) (1·10⁶ cells per injection). (F) Representative image showing the generated tumors per genotype. (G) Immunohistochemistry of SOX9 and Ki67 staining in SW620 derived tumors from pLKO or shSOX9 conditions (n = 4). (H) Quantification of percentage of Ki67 positive cells in SW620 derived tumors from pLKO or shSOX9 conditions (n = 4).