Figure 2

Palmitoylation contributes to Gαi2 coupling. (a) The colocalization between HA-tagged receptor and Gαi2 was determined in HEKOPRM1 and HEKC170A cells. Images were analyzed as described in Methods. (b) Anti-HA antibody was used to precipitate receptors in HEKOPRM1 and HEKC170A cells. Gαi2 precipitated with receptors was quantified and normalized to that in HEKOPRM1cells. (c) Co-immunoprecipitation was performed with Anti-Gαi2 antibody and the precipitated receptor was quantified. (d) CFPOPRM1 or CFPC170A was transfected into HEK cells with YFPGαi2. FRET analysis was performed. A two-tailed student t-test was used. Error bars and "*" represent the standard deviations and significant changes (p < 0.05, n > 3 for b and c; n > 10 for a and d), respectively. IP, immunoprecipitation; IB, immunoblotting.