Fig. 7
From: Super enhancers targeting ZBTB16 in osteogenesis protect against osteoporosis

BRD4 and RPAP2 promote ZBTB16 transcriptional pause release and elongation by synergistically regulating RPB1 CTD phosphorylation. a ChIP‒qPCR analysis showing the relative levels of pSer5 of the RPB1 CTD on ZBTB16 in NC and OB group MSCs. b ChIP‒qPCR analysis showing the relative levels of pSer2 of the RPB1 CTD on ZBTB16 in NC and OB group MSCs. c–e Western blot analysis of pSer5 and pSer2 in the cytoplasm, nucleus and chromatin (c). Scatter plots showing the relative levels of pSer5 (d) and pSer2 (e) in different protein extract fractions. f ChIP‒qPCR analysis showing the effects of RPAP2 knockdown, BRD4 knockdown and JQ1 treatment on the relative levels of pSer5 of the RPB1 CTD on ZBTB16 in MSCs. g ChIP‒qPCR analysis showing the effects of RPAP2 knockdown, BRD4 knockdown and JQ1 treatment on the relative levels of pSer2 of the RPB1 CTD on ZBTB16 in MSCs. h DNase-seq signal traces showing the accessibility of ZBTB16 chromatin at different time points during osteogenic differentiation. Shadows show the constituent SE enhancers targeting ZBTB16. i Dual-luciferase reporter assays showing the effects of RPAP2 knockdown, BRD4 knockdown and JQ1 treatment on the transcriptional activity of the constituent SE enhancers targeting ZBTB16 in MSCs. j Dual-luciferase reporter assays showing the effects of BRD4 and BRD4 ΔET overexpression on the transcriptional activity of the constituent SE enhancers targeting ZBTB16 in MSCs. The statistical data are represented as the means ± SEMs, n = 9, *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001