Fig. 1

Deacetylation of FOXP1 by HDAC7. a FOXP1 protein was enriched by immunoprecipitation (IP) from mesenchymal cells of human mesenchymal stem cells (MSCs) and was detected for acetylation by western blot with anti-Pan Acetyl-Lysine antibody (Ab). b A pcDNA-FOXP1-His plasmid was expressed in 293 T cells and FOXP1 was harvested via IP with anti-His Ab. Extensive acetylation was identified by mass spectrum analysis. c Mass spectral characterization of murine FOXP1 protein acetylation at amino acid T172. d Co-IP detection of the in vitro interaction of FOXP1 and HDAC7 in 293 T cells following their transfection with the indicated plasmids (representative of 3 independent experiments). e Co-IP detection of the in vivo interaction of FOXP1 and HDAC7 in bone marrow (BM) MSCs (representative of 3 independent experiments). f Immunostaining with anti-His (green) and anti-Flag (red) detects the co-localization of FOXP1 and HDAC7 within perinuclear region of C3H10T1/2 cells transfected with the indicated plasmids. Bar, 25 μm. g Deacetylation of FOXP1 by HDAC7 in 293 T cells. h Diagram depicting the expression subregions of FOXP1 protein. FOXP1-N: FOXP1 protein N-terminal(1-302aa); FOXP1-M: middle part (302-369aa), including zinc finger (ZF) and leucine zipper (LZ) domains; FOXP1-N/M: N-terminal and Middle part (1-369aa); FOXP1-C: C-terminal (369-673aa), containing the Forkhead (Fkh) domain [FOXP1-C(Fkh)]. Blue lines indicated the potential interaction domain between FOXP1 and HDAC7 protein. i–l Co-IP detection of the in vitro interaction of FOXP1-N/M and HDAC7-C in 293 T cells transfected with the indicated plasmids (representative of 3 independent experiments)