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. 2016 Jun 28;113(28):7786–7791. doi: 10.1073/pnas.1608061113

Fig. S5.

Fig. S5.

Biochemical interaction studies of Gpr161 and PKA. (A) GST pulldown analyzes of endogenous PKA subunits from HEK293 cell lysates in the presence or absence of 5 mM cAMP using GST and GST-CT variants. (B) cAMP-precipitation of endogenous PKA complexes and overexpressed Gpr161 variants tagged with Venus-YFP using two types of cAMP agarose. We have enriched PKA holoenzyme complexes using Rp-8-AHA-cAMP and activated PKA regulatory complexes using 8-AHA-cAMP. In the negative control experiment we added an excess of cAMP (5 mM) to the lysates to mask the cAMP binding sites in the R subunits for precipitation. An asterisk (*) indicates a degradation product of Gpr161-YFP.