Fig. 4

DPY30 promotes ZEB1 transcriptional expression through H3K4 methylation and regulates H3K4me3 binding to the promoter region of ZEB1 in CRC cells. A mRNA levels of EMT-related regulatory factors Snail1, Slug, Twist1, Twist2 and Zeb1 in CRC cells with DPY30 knockdown as measured by RT-qPCR. B The expression level of ZEB1 and H3 lysine methylation levels were assessed in CRC cells following DPY30 knockdown by Western blotting using whole-cell lysate; total H3 and β-actin were used as loading controls. C The intensity quantified statistical analysis of WB results. D Schematic presentation of the three regions relative to the ZEB1 transcriptional start site used as primers to test H3K4me3 occupancy. E ChIP-qPCR was performed to assess H3K4me3 occupancy at the ZEB1 transcriptional start site in HT29-shDPY30, SW480-shDPY30, and their control cells (E, left). IgG was used as a negative control (E, right). "Percentage of input" represents the ratio of DNA fragments of each promoter region bound by H3K4me3 to the total amount of input DNA fragments without H3K4me3 antibody pull-down. Data are expressed as mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001