Skip to main content
. 2016 May 26;7(25):37471–37486. doi: 10.18632/oncotarget.9650

Figure 6. Acetylation increases the DNA binding activity of Osx.

Figure 6

A. Evolutionary conservation analysis of the acetylation sites in Osx. Schematic representation of Osx, its functional domains, the two acetylated lysine residues that were mutated to arginines and a partial polypeptide sequence alignment of Osx with Sp1, Sp3 and Sp4. The highlighted boxes indicate the zinc-finger domain (from amino acid 309 to 376). The acetylated lysines are indicated by asterisks. B. Nuclear extracts (NE) from HEK 293T cells transfected with Flag-Osx expression plasmid alone or together with Flag-CBP-HA were incubated with a 32P-labeled Osx-binding oligonucleotide (MMP-13 probe) in the absence (lane 1 and 2) or presence of anti-Flag antibodies (lane 3). The reaction mixtures were analyzed by EMSA. Osx and CBP proteins were detected by western blotting with an anti-Flag antibody. C. Nuclear extracts from HEK 293T cells transfected with HA-Osx-GFP or HA-OsxK307R-K312R-GFP expression plasmids were incubated in reaction mixtures containing 32P-labeled MMP-13 probe, resolved by electrophoreses and visualized by autoradiography. Osx and OsxK307R-K312R proteins were detected by western blotting with an anti-HA antibody. Experiments were repeated at least three times.