Figure 6.

Modification of histone H3 by BIX01294 and TSA. Protein was isolated from bone marrow MSCs that were cultured in the absence or presence of BIX01294 and/or TSA. Following electrophoretic separation, the protein preparations were blotted with antibodies that recognized either total or specifically modified histone H3. (a), (b) Immunoblots of total protein demonstrated that methylation of H3K9 and H3K27 was reduced by G9a HMTase inhibition by BIX01294. (c) Treatment with the HDAC inhibitor TSA led to increased acetylation at H3K9. (d) While culturing MSCs with TSA also enhanced H3K27 acetylation, this increase was slight. However, the coaddition of BIX01294 to TSA synergistically stimulated the acetylation of this lysine residue. (e) Blotting for total histone H3 as control verified that each cellular sample consisted of equal amounts of protein.