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. 2005 Jan 3;102(2):349–354. doi: 10.1073/pnas.0408608102

Fig. 3.

Fig. 3.

The C subunit of PKA interacts with AKIP1. (A) Immunoprecipitation with the HA antibody showed that the HA-tagged C subunit specifically coprecipitated AKIP1a. Blots were probed with anti-C and anti-AKIP1 antibody. (B) GST pull down showed direct binding of the C subunit with GST-AKIP* as analyzed by SDS/PAGE. GST was used as control. (C) AKIP1a, AKIP1b, AKIP (residues 1–136), and H6-C subunit were in-vitro-transcribed, translated by using [35S]Met, separated by SDS/PAGE, and analyzed by autoradiography. Ten percent of the mix was loaded to obtain the total protein (Input). Pull down of the proteins with affinity matrix showed that the H6-C subunit interacted with AKIP1a, interacted weakly with AKIP1b, and failed to interact with AKIP (residues 1–136). The affinity matrix alone with AKIP1a and AKIP1b was used as control. (D) The C subunit and Δ(residues 1–14) C subunit were in-vitro-translated and pulled down with GST-AKIP* and GST, separated by SDS/PAGE, and autoradiographed. The C subunit and Δ(residues 1–14) C subunit interacted with GST-AKIP*.