Fig 1: Expression of KAT6B downstream molecules. qPCR was performed to measure the expression of RUNX2 (a), RUNX3 (b), COL2A1 (c), and COL10A1 (d) in primary chondrocytes from control (N = 10) and CS (N = 13) groups. e Western blotting was performed to measure the expression of RUNX2, RUNX3, COL2A1, and COL10A1 in primary chondrocytes from control and CS groups. CS, congenital scoliosis. *p < 0.05
Fig 2: DNA methylation analysis in blood samples from patients with congenital scoliosis. a A target region DNA methylation sequencing was performed to screen the differential methylation. Heat map of differential methylation in genes. b Principal component analysis (PCA) in congenital scoliosis samples and healthy samples. c The mean methylation levels and position of the top eight differentially expressed genes. CHST14, carbohydrate sulfotransferase 14; COL14A1, collagen type XIV alpha 1 chain; CREB3L2, cAMP responsive element binding protein 3 like 2; FGF23, fibroblast growth factor 23; INF2, inverted formin 2; KAT6B, lysine acetyltransferase 6B; ADAMTS7, a disintegrin and metalloproteinase with thrombospondin motifs 7; XYLT1, xylosyltransferase 1
Fig 3: Association of Cobb angles and methylation levels of eight genes, including CHST14, COL14A1, CREB3L2, FGF23, INF2, KAT6B, ADAMTS7 and XYLT1
Fig 4: KAT6B promotes chondrocyte proliferation. a qPCR was performed to measure the expression of KAT6B in primary chondrocytes from CS tissues after transfection. b Western blot was performed to measure the expression of KAT6B in primary chondrocytes from CS tissues after transfection. c EdU staining was performed to test cell proliferation in primary chondrocytes from CS tissues after transfection. DAPI, blue; EdU, red. Bar, 100 µm. d Western blotting was performed to measure the expression of RUNX2, COL10A1, ß-catenin, c-Myc, and VEGF in primary chondrocytes in primary chondrocytes from CS tissues after transfection. CS, congenital scoliosis. *p < 0.05
Fig 5: 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
Supplier Page from Abcam for Anti-KAT6B / MORF antibody