Fig. 2: High expression of TRIM24 correlates with activation of Wnt/β-catenin signaling pathway in colorectal cancer.

A The Gene set enrichment analysis (GSEA) based on the expression of TRIM24. CRC samples from TCGA were divided into two groups based on the median value of TRIM24 expression and then subjected to GSEA (https://rookieutopia.hiplot.com.cn/app_direct/BEST/) to investigate whether genes in the two groups were rich in meaningful biological processes. B GSEA showed that Wnt/β-catenin signaling and PI3K/AKT/mTOR signaling were hyper-activated in samples with high expression of TRIM24. NES represents normalized enrichment scores. C DLD1-WT/DLD1-KO-1 and SW620-WT/SW620-KO-1 cells were co-transfected with Renilla luciferase and TOPFlash plasmids (or FOPFlash plasmids, negative control). The cells were harvested 48 h after transfection, and firefly and Renilla luciferase substrates were added successively. The ratio of the detection value of firefly luciferase to the detection value of Renilla luciferase is taken to compare the difference in the results between the two groups of cells. Data were expressed as means ± SEM, two-tailed Student’s t-test. n  =  3 biological replicates per group. D Total RNA was extracted from DLD1-WT-1 and DLD1-KO-1 cells and mRNA profiles of MYC, CD44, CDC25A, EFNB2, HNF1A, and ACTB were analyzed by qRT-PCR. Data were expressed as means ± SEM, two-tailed Student’s t-test. n  =  3 biological replicates per group. E Whole-cell lysates from DLD1-WT/DLD1-KO-1 and SW620-WT/SW620-KO-1 cells were immunoblotted for indicated proteins, with Tubulin as a loading control. Shown are the representative results from three independent experiments.