Fig. 6: The dependency of acetylated SOX9 on SOX9 phosphorylation for Cyclin D1 gene induction and GMC proliferation in response to sublytic C5b-9.

Rat GMCs pretransfected with shSOX9 plasmid were treated with TSA (20 μM) for 30 min, followed by sublytic C5b-9 stimulation for 24 h or 3 h. GMC proliferation was determined by CCK-8 (a) and EdU incorporation assays (b). Cyclin D1 promoter activity and occupation by SOX9 were analyzed by luciferase reporter assay (c) and ChIP-PCR (d). The levels of SOX9 acetylation and phosphorylation and the protein expression of Cyclin D1 were determined by co-IP and IB (e). **p < 0.01 versus the shCTR + DMSO group; Δp < 0.05 or ΔΔp < 0.01 versus the shCTR + DMSO + sublytic C5b-9 group; ##p < 0.01 versus the shCTR + TSA + sublytic C5b-9 group. Data were represented as the means ± SD (n = 5 in each group for CCK‐8 assays, n = 3 in each group in the other experiments).