miR-23a knockdown increases MTSS1 protein levels and inhibits CRC cell and CCSC migration and invasion. A, crystal violet staining of a transwell migration assay of CCSC transduced with lentivirus containing anti-miR-23a shRNA or control sequences. B,C, anti-miR-23a or control sequence lentiviral transduced LoVo, SW480, and CCSC migration and invasion, 95% CI error bars are shown. For all comparisons p <0.002 by Student t test. D, miR-23a binding site in MTSS1 mRNA 3’UTR (nt 1949–1955) (top), bottom, miR-23a binding activity luciferase reporter assay in CCSC. CCSC were transfected with a plasmid containing the MTSS1 miR-23a binding site fused to the 3' UTR of Firefly luciferase and co-transfected with plasmids driving expression of pre-miR-23a or a control insert sequence. Luciferase protein levels and activity are suppressed when a miR binds specifically to the 3' UTR target sequence. E, MTSS1 protein levels in CCSC infected with lentivirus expressing a control or anti-miR-23a shRNA.