Fig. 5.

Circ_0002669 increases c-MYC transcriptional activity. A, B Relative expression of MYCBP and c-MYC downstream genes was determined, by qRT-PCR and western blotting, in OS cells transfected with MYCBP plasmid or siRNAs. C, D Relative expression of MYCBP and c-MYC downstream genes was determined in circ_0002669-overexpressing or -knockdown OS cells by qRT-PCR and western blotting. E CHIP-qPCR showing circ_002669 enhanced c-MYC occupancy on the c-Jun, CCND1 and CDK4 promoters in U2OS cells. F Circ_002669 in U2OS cells increased transcription of the c-Jun promoter. G MYCBP expression in OS (n = 12) and normal tissues (n = 5) was determined by qRT-PCR. H MYCBP expression in OS and normal tissues (n = 7) was determined by western blotting. I MYCBP expression in OS tissues (n = 72) and normal tissues (n = 6) was determined by IHC. J MYCBP expression in control or circ_002669-overexpressing samples derived from xenografts (scale bar, 100 μm). K Pearson correlation analysis showing the correlation between expression of MYCBP and circ_002669 in OS tissues. Data shown are from three independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001