Fig. 1. Scheme and protein expression levels of wild-type and SCA14 mutant PKC. (A) Schematic representation of the primary structure of PKC, including the V1-domain containing the pseudoinhibitory substrate, the regulatory domain comprising the C1 and C2 domains and the catalytic domain comprising the C3 and C4 domains. SCA14 mutations used in this study are indicated in black. (B) Schematic representation of the constructs. Both full-length PKC and the C1B subdomain are fused to GFP and RFP. (C) Western blot for PKC, demonstrating equal expression of the different PKC alleles (left). Similar expression levels of the C1B fusion proteins were also detected using anti-GFP (right). Actin was used as a loading control. (D) Confocal images of transiently transfected HeLa cells with either full-length PKC-GFP or C1B-GFP fusion plasmid. SCA14 mutations do not alter the cellular localization and distribution of full-length PKC and C1B proteins in HeLa cells. Scale bars: 10 µm.