Fig. 6

CircDNAJC11 modulates MAPK6 expression via TAF15. A Silver staining plot of proteins in MCF-7 cells was pulled down from the biotin-labeled circDNAJC11 probes and the control probes. B TAF15 peptide was determined by mass spectrometry analysis. C Binding of circDNAJC11 to TAF15 was determined by RIP assay using an anti-TAF15 antibody and an IgG control antibody. D FISH and IF double staining showing the localization of circDNAJC11 (red) and TAF15 (green) in MCF-7 and SK-BR-3 cells. Scale bar, 50 μm. E After BC cells were transfected with corresponding siRNAs for 48 h, the expression levels of the MAPK6 signaling pathway-associated proteins in BC cells of each group were evaluated by western blot. F The mRNA expression levels of MAPK6 in MCF-7 and SK-BR-3 cells transfected with si#NC or siRNAs for circDNAJC11 were assessed by qRT-PCR. G RIP experiments confirmed the binding of TAF15 protein and MAPK6 mRNA. H–K The effect of knockdown or overexpression of circDNAJC11 on MAPK6 mRNA stability in the presence of transcription inhibition using ActD (4 μg/mL) was determined by qRT-PCR. For F, β-actin was utilized as a loading control. Data were presented as mean ± SD and representative of three independent experiments in C, F, and G–K. C, G, H, and J were analyzed by Student’s-t test, and F, I, and K were analyzed by ANOVA. * P < 0.05, ** P < 0.01, ***P < 0.001, ns, no significance