A-C, Constructs containing Cpne5, 8 or 9 genes were transfected into the HEK293-Cre cells. Localization of the two fusion proteins is revealed by immunostaining for GFP (green, first column), HA (red, second column) and Cpne (white, third column) antibodies indicate their localization in the HEK293Cre cells. The meGFP is localized to the cell membrane in all the transfected cells. A’-C’’’, The dashed boxes in A, B and C are zoomed in to show the differences in sub-cellular distribution between Cpne5, 8 and 9, respectively. HA immunostaining indicates the presence of Copines mostly in the cytosol and an absence in the nuclei. Pan Cpne 5/8/9 antibody however has some localization in the nuclei of the cell (C, C’’’). n= 4 each. Scale bar in A: 50μm.