Fig. 2

Sec23a inhibits the stemness of M14-SE cells. a, b. RT-qPCR and Western blot were performed to measure Sec23a expression in M14-Parental and M14-SE cells. c, d RT-qPCR and Western blot were performed to confirm effective gene manipulation of Sec23a expression in M14-SE cells. e Analysis of mRNA expression of Aldh1, CD133, Nanog, Oct4 and Sox2 in M14-SE cells with Sec23a interference or overexpression. GAPDH expression was used as a reference control. f–h Clonogenic spheroid formation rate, morphology and average spheroid diameter of M14-SE cells with Sec23a interference or overexpression in the single-cell cloning assay. i Serial spheroid formation assay of M14-SE cells with Sec23a interference or overexpression, bar = 200 µm. j Tumors resected from nude mice inoculated with M14-SE cells with Sec23a interference or overexpression, bar = 1 cm. (k and l). Tumor volume and weight, *P < 0.05, **P < 0.01, ***P < 0.001