Fig. 6: TRIM11 and KDM5C regulate cell migration through MCAM enhancer.

A Heatmap of genes up-regulated in KDM5C-KD cell lines and rescued in DKD cell lines. Genes related to cell migration are highlighted at right. B The UCSC browser view to show KDM5C, H3K27ac and H3K4me3 enrichment on MCAM enhancer. C MCAM mRNA expression level in TRIM11-KD, KDM5C-KD and DKD cell lines (n = 3). D H3K27ac and H3K4me3 enrichment on MCAM enhancer in KDM5C-KD and DKD cell lines, analyzed by ChIP-qPCR (n = 3). E ChIP-qPCR analysis of KDM5C occupancy on the MCAM enhancer. IgG was used as the control. Statistics were presented as mean ± SD. F CRISPR-Cas9-KRAB system and two different sgRNAs were used and stable cell lines were established in MDA-MB-231. MCAM expression was detected with quantitative RT-PCR. G MCAM was stably knocked down in KDM5C knockdown MDA-MB-231 cells using the CRISPR/Cas9 system. H Cell migration of the cell lines in (G) were tested by transwell assay. Four views of the cell migration images were taken for each cell line. The number of moving cells was counted by image J. Relative cell migration was shown in the figure. I Quantitative RT-PCR analysis of Mcam gene in breast tumors of Trim11^+/+ and Trim11^+/− MMTV-PyVT mice (n = 5, per group, 3 replicates). J Overall survival was analyzed and plotted using the Kaplan–Meier method. The survival rates for patients with high and low MCAM expression were plotted as red and blue lines, respectively. The number of patients in each group was shown in parentheses, p values were calculated using a log-rank test. Histograms were presented as mean ± SD (C, D, E, G) or SEM(H). ^*p value ≤ 0.05, ^**p value ≤ 0.01, ^***p value ≤ 0.001.