Figure 5
The double helical motif of OCIAD2 is necessary for interaction with OCIAD1. (A) Schematic representation of full-length (FL), N-terminal (N), hydrophobic region (Hph) and C-terminal (C) fragments of OCIAD2 generated for this study. Numbers indicate amino acid positions. Hx: predicted alpha helix. (B–H) Confocal images of HEK293 cells transfected with OCIAD2 reporter constructs, showing localization of ectopic OCIAD2-reporter with endogenous OCIAD2 (B–D) or endogenous OCIAD1 (E–H) as indicated, detected by fluorescence immunostaining with respective antibodies. Insets show magnified view of the boxed region. Co-localization plots are to the right of each panel. Scale bar: (B–D): 10 µm; (E–H): 5 µm. (I–O) Cell lysates from untransfected HEK293 cells (I) or those expressing various OCIAD2 (J, K, L, O) or OCIAD1/Asrij (M,N) constructs as indicated on the top of each panel, were subjected to immunoprecipitation (IP) followed by immunoblotting (IB) with antibodies as indicated. Protein size markers (in kDa) are indicated to the left. At least two independent immunoprecipitation experiments with two technical replicates each were performed and similar results were obtained. Full-length blots are presented in Supplementary Figure S6.
