Fig. 4

IPW upregulates miR-29c expression by enhancing H3K4 trimethylation. A Experimental scheme of ChIP assay. B ChIP assay was performed in DCIS-control and DCIS-IPW cells using specific antibodies to H3K4me3- (left panel) and H3K27me3- (right panel). The enrichment of miR-29c promoter region was examined by qRTPCR (n = 3/group). C Experimental scheme of SET methyl transferase immunoprecipitation assay. D DCIS (control) and DCIS-IPW cells were immunoprecipitated by IgG or SET-methyltransferase antibody followed by isolation of RNA and cDNA synthesis. Enrichment of IPW was examined by qRT-PCR (n = 3/group). E DNA was isolated from DCIS-control and DCIS-IPW cells immunoprecipitated with IgG or SET methyltransferase (as shown in C) followed by quantitation of miR-29c promoter enrichment by qRT-PCR (n = 3/group) F MCF10A cells were immunoprecipitated by IgG or SET methyltransferase antibody followed by isolation of bound RNAs. Enrichment of IPW in the IgG- or SET methyltransferase-bound RNAs was examined by qRT-PCR. G MCF10A cells were immunoprecipitated by IgG or SET methyltransferase antibody followed by isolation of bound DNAs. Enrichment of miR-29c promoter region in the IgG- or SET methyltransferase-bound DNAs was examined by qRT-PCR. Statistical inference between two groups was determined by unpaired two-tailed Student’s t test and data are represented as mean + S.E.M. H Hypothesis figure showing IPW interaction with SET methyltransferase and locating the later to miR29c promoter region. This results in enhanced H3K4me3 leading to increased miR-29c expression. (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). ns: not significant