Skip to main content
. 2002 Nov;22(22):7907–7918. doi: 10.1128/MCB.22.22.7907-7918.2002

FIG. 3.

FIG. 3.

Coimmunoprecipitation of acetyltransferases and PyLT. (A) Analysis of PyLT coimmunoprecipitated with Gal4 fusion PCAF. Cell extracts from NIH 3T3 cells transiently cotransfected with expression vector for PyLT and an expression vector for Gal4 fusion acetyltransferase were precipitated with rabbit polyclonal anti-GalDB antibody. One half of the immune complexes (top band), equivalent to one half of whole-cell extracts (WCE), and one fiftieth of WCE for measurement of input PyLT (lower band) were analyzed by Western blots. (B) Analysis of PyLT coimmunoprecipitated with Gal4 fusion GCN5 as described for panel A. (C and D) Analysis of acetyltransferases coimmunoprecipitated with PyLT. Cell extracts from NIH 3T3 cells transiently cotransfected with expression vector for PyLT and an expression vector for Gal4 fusion acetyltransferase were incubated overnight at 4°C with mouse monoclonal anti-PyLT antibody and mouse normal serum immunoglobulin G (IgG). One half of the immune complexes, equivalent to one half of WCE, and one fortieth of WCE for measurement of input Gal4 fusion acetyltransferases were analyzed by Western blotting by using rabbit polyclonal anti-GalDB antibody (top lane) and one fiftieth of WCE for measurement of input PyLT analyzed by Western blotting by using anti-PyLT antisera (lower lane). Gal4 fusion acetyltransferases transfected and antibodies (anti-PyLT and normal IgG) used for immunoprecipitation are indicated above each lane. α PyLT, anti-PyLT antibody; α GalDB, anti-GalDB antibody; IP, immunoprecipitation.