Fig. 7.
Phosphorylation of overexpressed hTRPC1 and mTRPC6 using CAT of cAMP-PK.
500 μg membrane proteins from QBI-293A cells overexpressing either hTRPC1 or mTRPC6 were incubated with γ[32P] ATP in the absence of CAT (CON), presence of 350 U CAT (+PKA CAT), or presence of 1 μM cAMP-PK inhibitor peptide (+PKA INHIB). (A) Autoradiography of Ank immunoprecipitations of overexpressed hTRPC1 and subsequent probing of the Western blot with the Ank antibody (5 μg/mL). (B) Autoradiography of anti-TRPC6 immunoprecipitations of overexpressed mTRPC6 and probing with anti-TRPC6 antibody (1/400). Note both partial and fully glycosylated forms of mTRPC6 undergo phosphorylation. Molecular size markers are noted in kilodaltons (kDa) and the TRPC protein identifications indicated with arrows.