Distribution of Gαi subunits in AtT-20 cells stably expressing calnuc-GFP or calnuc(ΔSS)-CFP. (A-F). Deconvoluting analysis of cells expressing calnuc-GFP which had been doubly stained for ACTH and endogenous Gαi1/2 or Gαi3. Both Gi subunits are associated with RSG containing ACTH (yellow in C and F). (G-L). Deconvolution analysis of cells expressing calnuc-GFP and doubly stained for GFP and either Gαi3 or Gαi1/2 showing that the Gi subunits are distributed on the PM (arrows) and partially colocalize with calnuc-GFP (yellow in I and L) which is distributed in an interrupted pattern on the PM. Calnuc-GFP also colocalizes with Gαi3 on the Golgi (G). (M-O). Similar deconvolution analysis of AtT-20 cells expressing calnuc(ΔSS)-CFP showing that some of the Gαi1/2 colocalizes with cytosolic targeted calnuc(ΔSS)-CFP on RSG (yellow dots in O) and the PM (arrows). Bar = 10 μm.