Fig. 5

MILIP links YBX1 to translational activation of Snai1 a YBX1 was co-pulled down with Snai1 mRNA in Caki-1 and ACHN cells as shown in RNA pulldown (RPD) assays. GAPDH was included as a negative control. S: scramble; AS: antisense. b Snai1 mRNA was co-precipitated with YBX1 as detected by RNA immunoprecipitation (RIP) assays. GAPDH protein was used as a negative control. c Induced knockdown of MILIP decreased the amount of Snai1 mRNA associated with YBX1 protein as shown in RIP assays. Two-tailed Student’s t-test. d, e SiRNA knockdown of YBX1 diminished the increase of Snai1, N-cadherin, MMP2 and Vimentin and the decrease of E-cadherin caused by MILIP overexpression in 769-P cells. f, g SiRNA knockdown of YBX1 diminished the increase of cell migration and invasion caused by MILIP overexpression in 769-P cells. Relative area of migrated and invasive cells was quantitated using the ImageJ software (g). One-way ANOVA followed by Tukey’s multiple comparisons test. Data are representatives or mean ± s.d.; n = 3 independent experiments