Fig. 7

CircMYO10 promotes H4K16Ac at the promoter region of C-myc via miR-370-3p/RUVBL1 axis. a The occupancy of the MYC promoter by RUVBL1, LEF1, TIP60. a-f Chromatin was collected from MG63 and U2OS cells and was subjected to immunoprecipitation with IgG or anti-LEF1-antibodies or anti-TIP60 antibodies or anti-RUVBL1 antibodies or anti-histone H4K16Ac. Recovered DNA was used as a template in PCR amplification using the pair of primers flanking the LEF1-binding site at − 1804. 2% Input was used as a control in the detection of DNA immunoprecipitated with RUVBL1, LEF1 and TIP60 with 0.5% input for DNA immunoprecipitated with H4K16Ac. b-c The recruitment of RUVBL1 to the promoter region and measurement of H4K16Ac by CHIP assays in MG63 cells stably transfected with either b shRUVBL1 or b TIP49D302N or c RUVBL1 or c shCTNNB1 or c shTIP60. d MG63 cells stably transfected with pre-miR-370-3p inhibited the recruitment of RUVBL1 to the promoter region and H4K16Ac, both of which was restored by co-transfection with RUVBL1. e MiR-370-3p sponge promoted the recruitment of RUVBL1 and H4K16Ac which was abrogated by shRUVBL1. f Recruitment of RUVBL1 to the promoter region was suppressed upon transfection with shcircMYO10 but partially restored by co-transfection with either miR-370-3p sponge or RUVBL1 in MG63 cells. The acetylation of H4K16 was highly correlated with the recruitment of RUVBL1. b-f Data represents the mean ± SD (n = 3). Three independent assays were performed in the above assays. b-f * P < 0.05, ** P < 0.01, *** P < 0.001 (Student’s t-test)