Skip to main content
. 2010 Nov 21;3(2):121–132.

Figure 6.

Figure 6

The interaction between CtBP and KLF8 is negatively regulated by PCAF and p300 mediated acetylation. (A) PCAF, p300, or HDAC inhibitors inhibit KLF8 interaction with CtBP. HEK293 cells were co-transfected with expression vectors coding for HA-KLF8, PCAF, or p300 or treated with HDAC inhibitors (trichostatin A, sodium butyrate, and nicotinamide) as indicated. Endogenous CtBP was immunoprecipi-tated (IP) using an anti-CtBP antibody, and blotted (IB) with either anti-HA (panel a), or anti-CtBP antibodies (panel b). Conversely, KLF8 was immunopre-cipitated using anti-HA, and blotted using anti-CtBP (panel c) or anti-HA antibodies (panel d). (B) Alignment of KLF8 orthologs shows that the PVDLS motif and lysines 93 and 95 are highly conserved. (C & D), Acetylation at lysines 93 and 95 inhibits KLF8 interaction with CtBP. Experiments were performed similarly as in A except that K93R or K93Q (C) or K95R or K95Q (D) was transfected. Shown are representative experiments where n=3. Expression of the transfected constructs was verified by whole cell lysate blotting (data not shown).