(A) SW48 derivative cell lines were treated with indicated doses of MMC for 3 days and cell numbers were determined by CCK8 assay. (B - C) U251MG derivative cell lines were treated with indicated doses of MMC for 3 days, and cell numbers were determined by CCK8 assay. (D-E) Derivatives of (D) SW48 and (E) U251MG cell lines were treated with indicated doses of MMC for 24 hours, and clonogenic formation assays were performed to determine the relative colony formation. (F) Derivative U251MG and SW48 cell lines (shCtrl or shPRMT5) were subcutaneously implanted in nude mice and tumors were treated with MMC (1 mg/kg via intraperitoneal injection once per week for 3-5 weeks), and tumor sizes were measured twice per week starting from the first treatment, and tumor size’s doubling times were determined and calculated (from the left to the right group, n=5, 6, 4, 7 (for U251MG) and n=6, 7, 6, and 8 (for SW48), respectively). n.s. not significant; *p<0.05; ** p<0.01.