Figure 4.

KLF8 sumoylation is negatively regulated by acetylation at lysines 93 & 95. (A) HEK293 cells were co-transfected with expression vectors coding for Myc -SUMO-1, HA-KLF8, or HA-mutant KLF8 (KàR or KàQ). Lysates were collected with NP-40 lysis buffer supplemented with 20 μM of the desumoylase inhibitor NEM followed by immunoprecipitation (IP) using an anti-HA antibody, and western blotting (IB) with either anti-Myc or anti-HA antibodies. Expression of the transfected constructs was verified by whole cell lysate blotting (data not shown). (B) The expression of SUMOylated KLF8 relative to total expression of KLF8 that is normalized to 1. Results represent mean ±S.E. where n=3.