Fig. 4

MILIP forms an RNA-RNA duplex with Snai1 mRNA to promote Snai1 translation a In vitro-synthesized MILIP bound to in vitro-transcribed Snai1 mRNA as shown in domain-specific chromatin isolation by RNA purification (dChIRP) assays. This binding was abolished when the MILIP-BR at Snai1 mRNA or the DFO within MILIP were deleted (Snai1-ΔMILIP-BR and MILIP-ΔDFO, respectively). BR, binding region. DFO, duplex-forming oligonucleotides. b Endogenous MILIP co-precipitated the endogenous Snai1 mRNA in Caki-1 and ACHN cells as shown in dChIRP assays. GAPDH protein was included as a negative control. S: scramble; AS: antisense. c-e Overexpression of wild-type MILIP but not MILIP with DFO deleted increased Snai1 protein expression levels (c) and cell migration and invasion (d, e) in 769-P cells. Relative area of migrated and invasive cells was quantitated using the ImageJ software. Scale bar, 1 mm. One-way ANOVA followed by Tukey’s multiple comparisons test. Data are representatives or mean ± s.d.; n = 3 independent experiments