Fig. 6
From: An intact helical domain is required for Gα14 to stimulate phospholipase Cβ
Ability of different chimeras to interact with AC and TPR1. a HEK293 cells were transiently transfected with the wild-type or constitutively active mutants of Gα protein and chimeras indicated in the figure. The transfectants were labelled with [3H]adenine (1 μCi/ml) in 1 % FBS/MEM. The labelled cells were treated with 50 μM of FSK for 30 min before subjected to cAMP accumulation assay. cAMP fold inhibition was calculated as the ratios of QL-induced to wild-type cAMP inhibition. Data represent the mean ± S.E.M. of three independent experiments, n = 3. *, cAMP accumulation was significantly inhibited as compared to corresponding wild-type transfected cells; Dunnett t test, p < 0.05. b HEK293 cells were transiently co-transfected with FLAG-TPR1 in combination with Gα proteins and the indicated chimeras. Cell lysates were immunoprecipitated by anti-FLAG affinity agarose gel. The immunoprecipitates were immunoblotted with anti-Gα14, anti-Gαz or anti-FLAG antiserum. Crude lysates were used to examine the expression levels of Gα14, Gαz, Gα14/Gαz chimeras or FLAG-TPR1 by Western blot analysis. The immunoblots shown represent one of three sets of immunoblots; two other sets yielded similar results