Interaction of Pex26p with Pex14p and Pex5p.
A, immunoprecipitation assays of FLAG-Pex26p with Pex14p and Pex5p. FLAG-PEX26 or PEX26 was transfected to pex14 ZP161 cells together with or without PEX14 (lanes 1–6), and lysates were subjected to immunoprecipitation with antibody against FLAG. Input, 5% input used for immunoprecipitation (IP). Immunoprecipitated proteins were analyzed by SDS-PAGE and immunoblotting using antibodies to human Pex26pC (upper panel) and rat Pex14pC (middle panel). Endogenous Pex5p was detected with anti-HsPex5pN antibody (lower panel). B, Pex14p and Pex5p are binding partners of Pex26p. Lysates from HEK293 cells (2 × 106) were subjected to immunoprecipitation with pre-immune and antisera against Pex26pC (lanes 2 and 3). One-eighth aliquot of the input used for IP was loaded in lane 1. Immunoprecipitates were analyzed by immunoblotting with antibodies against Pex26pC, Pex14pC, Pex5pN, Pex1pC, and Pex6pN.