Figure 3
From: DNA polymerase ι is acetylated in response to SN2 alkylating agents
In vivo and in vitro acetylation of Polι by p300. (a) Immunoprecipitation assay of extracts from HEK293T cells co-transfected with FLAG-Polι and wild-type p300-c-myc. As indicated, prior to harvesting, cells were treated either with DMSO or with deacetylase inhibitor (DKAc) cocktail. (b) Immunoprecipitation assay used extracts from HEK293T cells co-transfected with FLAG-Polι and wild-type p300-c-myc, or FLAG-Polι and p300 containing the D1399Y mutation that abolishes the acetyltransferase activity of p300. 24hrs prior to harvesting, cells were treated with DKAc inhibitor. In panels (a,b), Polι acetylation was verified by western blot using antibodies against acetylated lysines (KAc). Polι was visualized using polyclonal rabbit antibodies against a C-terminal fragment of Polι. (c) In vitro acetylation of recombinant His-tagged Polι by p300. The reaction products were resolved by SDS-PAGE and analyzed by western blot using antibodies against KAc and against Polι.
