Fig. 6: Formation of snail/PRMT5/NuRD(MTA1) repression complex on transcriptional targets.

A, B Clones in which Snail, PRMT5, and MTA1 stably knocked down were compared to the parental cell line with respect to mRNA and protein levels of indicated genes in HeLa cells. The mRNA levels were normalized to those of GAPDH and β-actin served as a loading control for western blotting. Error bars represent the mean ± SD of three independent experiments. (*p < 0.05, **p < 0.01, ***p < 0.001, and two-tailed unpaired t-test). C Equal amounts of genomic DNA from HeLa cells were used for dot blot assays with antibodies against 5mC or 5hmC. D ChIP and Re-ChIP experiments were performed in HeLa cells with the indicated antibodies. E HeLa cells were infected with lentiviruses carrying the indicated shRNAs. qChIP analysis of the selected promoters was performed using antibodies against Snail, PRMT5, MTA1, Mi-2, H4R3me2s, H3R8me2s, or H3Ac. H3 was detected as an internal control. Results are represented as the fold-change over the control with GAPDH as a negative control. F, G HeLa cells were transfected with the indicated specific shRNAs or/and expression constructs for cell invasion assays. The invaded cells were stained and counted. The images represent one field under microscopy in each group. The efficiency of protein knockdown or overexpression was verified by western blotting. F-∆PRMT5, PRMT5 expression construct without the SAMD domain (enzymatic domain). H Analysis of public datasets (GSE68339 and GSE72723) for the expression of PRMT5 and E-cadherin or TET1 in cervical cancer. The relative levels of E-cadherin and TET1were plotted against that of PRMT5.