Fig. 4
MIR100HG and hnRNPA2B1 collaboratively regulate TCF7L2 mRNA stability and activate Wnt signaling. a Upset plot showing the number of significantly up- or down-regulated genes in multiple comparisons of the RNA-sequencing data of HCT116 cells after MIR100HG or hnRNPAB1 knockdown. b Immunoblots and qPCR analyses of TCF7L2 expression in CC and CC-CR cells after manipulation of MIR100HG (left) or hnRNPA2B1 (right) expression. Representative of three independent experiments. c, d Assessment of TCF7L2 mRNA half-life (t1/2) in hnRNPA2B1- (c) or MIR100HG- (d) silenced CC-CR cells. n = 3 independent biological replicates. e Assessment of TCF7L2 mRNA half-life in MIR100HG-overexprssing CC and Caco-2 cells after hnRNPA2B1 knockdown. n = 3 independent biological replicates. f Colony counts for 3D-cultured MIR100HGKOE4 (left) or hnRNPA2B1-silenced (right) CC-CR cells after TCF7L2 overexpression in the presence or absence of CTX (3 μg/ml), n = 3 independent experiments performed in triplicate. g Extent of migration and invasion using Transwell migration and invasion assays for MIR100HGKOE4 (left) or hnRNPA2B1-silenced (right) HCT116 cells after TCF7L2 overexpression. n = 3 independent biological replicates. h Immunoblots of E-cadherin and vimentin in CC-CR and LoVo cells after the indicated treatment. Representative of three independent experiments. i Top/Fop flash luciferase reporter activity in HEK293T cells after indicated treatment. n = 3 independent biological replicates. j qPCR analyses of Wnt target genes in CC and CC-CR cells after manipulation of MIR100HG (j) or hnRNPA2B1 (k) expression. n = 3 independent biological replicates. **P < 0.01, *P < 0.05. Data represent mean ± s.d., n.s., not significant