Characterization of AtT-20 cells stably overexpressing calnuc-GFP or Calnuc(ΔSS)-CFP. (A). Immunofluorescence showing that most of the endogenous calnuc is found in the Golgi region (G) with some found on the PM (arrow) (B) Stably expressed calnuc-GFP in live AtT-20 cells is also concentrated in the Golgi with some found on the PM, but it is also associated with RSG stored at the tips of cell processes. (C) Stably expressed mutant calnuc(ΔSS)-CFP is distributed throughout the cytoplasm in live AtT-20 cells. (D-F). Deconvolution analysis of immunofluorescence images demonstrates that calnuc-GFP partially colocalizes with ACTH in RSG (yellow in F) at the tips of the cell processes (arrowhead in F) and in the Golgi region (G). (G-I). Deconvolution analysis of AtT-20 cells stably expressing calnuc(ΔSS)-CFP demonstrates that cytosolic calnuc is found on RSG and partially colocalizes with ACTH (yellow in I).). Bar = 10 μm.