Fig. 3

EZH2 mediates trimethylation of H3K27 on KAT6B promoter. a Immunohistochemical staining for KAT6B in facet joint tissues. SCB, subchondral bone; DZ, deep zone; SZ, superficial zone. Bar, 50 μm. b Western blot was performed to measure the expression of KAT6B in primary chondrocytes (upper), and quantification of the bands (lower). c qPCR was performed to measure the expression of KAT6B in primary chondrocytes from control (N = 10) and CS (congenital scoliosis, N = 13) groups. d ChIP was performed in primary chondrocytes from control and CS (congenital scoliosis) using the indicated antibodies or IgG, and qRT–PCR was performed to test the promotor of KAT6B. e Schematic diagrams of the KAT6B promoter segments and mutations. Numbers indicate the nucleotides relative to KAT6B TSS (transcriptional start site) (upper panels). Luciferase reporter activities of human 293T cells transiently co-transfected with luciferase reporter constructs containing the wild-type sequence of KAT6B promoter or its mutant counterparts, together with EZH2 for 48 h