Fig 2: LncRNA-DANCR regulates Wnt/ß-catenin signaling. (A) ß-Catenin activity in A549 cells cotransfected with TOPFlash luciferase reporter + DANCR shRNAs compared to cells transfected with TOPFlash luciferase reporter + scramble control (n = 3, t test, *** p < 0.001, **** p < 0.0001); (B) fold change of ß-catenin protein expression in A549 DANCR KD cells compared to scramble control (n = 3, t test, * p < 0.05); (C) fold change of CMYC and AXIN2 gene expression in A549 DANCR KD cells compared to scramble control (n = 3, t test, * p < 0.05, ** p < 0.01, *** p < 0.001); (D) fold change of c-myc and Axin2 protein expression in A549 DANCR KD cells compared to scramble control (n = 3, t test, * p < 0.05, ** p < 0.01).

Fig 3: SLCO4A1-AS1 activates Wnt/ß-catenin signaling in CRC. a Gene Set Enrichment Analysis (GSEA) showed that SLCO4A1-AS1 highly expression was positively related to cell division and G1/G2 phase transition. Samples in Li’s cohort (GSE104836) were divided into 2 groups based on expression of SLCO4A1-AS1. NES, normalized enrichment score. b GSEA showed that SLCO4A1-AS1 expression was positively correlated with canonical Wnt signaling according to Li’s cohort (GSE104836). c Expression heatmap of the target genes of Wnt/ß-catenin signaling showed that more activation of Wnt/ß-catenin signaling appeared in SLCO4A1-AS1high group. d, e Real-time PCR confirmed that SLCO4A1-AS1 knockdown significantly downregulated Wnt/ß-catenin signaling in HCT116 and SW480 cells. f The protein levels of AXIN2, LGR5 and MYC in HCT116 and SW480 cells after SLCO4A1-AS1 silence were measured by western blot (WB). g-j The mRNA levels of AXIN2, LGR5, SOX4 and MYC were positively correlated with the expression of SLCO4A1-AS1 in the 50 CRC samples. qRT-PCR was performed and GAPDH acted as loading control. *p < 0.05 and **p < 0.01

Fig 4: HOXC6 contributes to activation of Wnt/ß-catenin pathway and EMT.A Pairwise correlation between Axin2 and HOXC6 using TCGA expression profiles. B Pairwise correlation between RNF43 and HOXC6 using TCGA expression profiles. C Expression of ß-catenin, c-Jun, RNF43, Axin2, DKK1, and HOXC6 was evaluated by WB in whole-cell lysates of HOXC6-OE, HOXC6-KD, and their control groups in the HCT116 cell line. D The expression of ß-catenin, c-Jun, and HOXC6 was evaluated in the cytoplasm and nucleus of HCT116 cells in the HOXC6-OE, HOXC6-KD, and control groups. Tubulin was used as a cell plasma protein and whole-cell protein loading control. Histone-H3 was used as a nuclear protein loading control. E GSEA showed that the EMT process was significantly enriched in the HOXC6-overexpressing group using TCGA dataset. F Protein expression of ZO-1, E-cadherin, vimentin, snail, and HOXC6 in three different treated HCT116 groups. G Relative mRNA expression of ß-catenin, DKK1, and HOXC6 in the NC, HOXC6-KD, and HOXC6-OE groups in HCT116 cells. *: P < 0.05; ***: P < 0.001.