Fig. 2. CBX4 increases Runx2 via recruiting GCN5 to the Runx2 promoter in osteosarcoma cells.

a, b The indicated stable cells transfected with the Runx2-Luc reporter for 48 h were subjected to the luciferase activity assay as described in Methods section. c The ChIP assay was performed in U2OS cells using anti-CBX4 antibody or IgG antibody, as indicated, of three independent experiments. The p16 promoter was used as the positive control. d, e ChIP-qPCR analysis of the occupancies of CBX4, GCN5, H3K27Ac, and Pol II on the Runx2 promoter in the indicated stable cells. f U2OS/MTX300 cells were subjected to immunoprecipitation (IP) using anti-CBX4 antibody or anti-IgG antibody followed by Western blotting as indicated of three independent experiments. g, h U2OS cells expressing pSIN-Vector or pSIN-CBX4 were transiently transfected with siGCN5 as indicated and then analyzed by qRT-PCR (g) and Western blotting (h). i, j The indicated stable cells co-transfected with the Runx2-Luc reporter and HA-GCN5 for 48 h were subjected to the luciferase activity assay as described in Methods section. k–m The indicated U2OS/MTX300 cells were subjected to Western blotting, cell migration and invasion assays. The bars indicate the SD. The results are expressed as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 using the two-sided Student’s t-test. n.s no significance. Source data are provided as a Source Data file.