Figure 3

Runx2 recruitment on BMP-3B gene promoter and interaction with Suv39h1. (a) Lung cancer cells (H-1299) stably expressing wild type Runx2 (left panel) or shRNA-Runx2 (right panel) were subjected to chromatin immunoprecipitation assay with Runx2, Suv39h1, H3K9 or control IgG-G (goat) and IgG-R (rabbit) antibodies. Immunoprecipitated DNA samples were further amplified by real-time qPCR with primer pairs for proximal promoter (-1.0kb) of BMP-3B gene. A schematic diagram above indicates Runx binding motifs (solid bars) on BMP-3B promoter. The BMP-3B ChIP primer pairs are indicated by arrows in the schematic diagram. (b) Co-immunoprecipitation assay shows Runx2 interaction with Suv39h1 in H-1299 lung cancer cells. Whole cell extracts from lung cancer H-1299 cells were incubated with Runx2, Suv39h1 or control IgG antibodies. Western blots showing Runx2 and Suv39h1 proteins as detected by αRunx2 and αSuv39h1 antibodies.