Fig. 6

Validation of the transcriptional regulation of SMYD3, MTA1, and MTA2 on target genes. a A heatmap presents the selected target genes that were coregulated by SMYD3, MTA1, and MTA2 in the RNA-seq analysis. b LM3 cells were infected with control siRNA, SMYD3 siRNA, MTA1 siRNA, or MTA2 siRNA. RT–PCR analyses of partial transcriptional target expression in LM3 cells infected with the indicated siRNAs are shown. c qChIP analysis of the selected promoters was performed using antibodies against IgG, SMYD3, MTA1, and MTA2. Results are represented as the fold change over IgG with glyceraldehyde 3-phosphate dehydrogenase as a negative control. 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). d ChIP/Re-ChIP analysis of the IGFBP4 promoter was performed using antibodies against SMYD3, MTA1, MTA2, HDAC1, HDAC2, and RBBP4. IgG served as a negative control. e qChIP analysis of the recruitment of the indicated proteins on the IGFBP4 promoter in LM3 cells after infection with control siRNA, SMYD3 siRNA, MTA1 siRNA, or MTA2 siRNA. Purified rabbit IgG was used as a negative control. Error bars represent the mean ± SD of three independent experiments. *P < 0.05 and **P < 0.01 (two-tailed t-test). f Western blot analyses of IGFBP4 protein expression in LM3 cells infected with the indicated siRNAs are shown