Figure 2. RPGR interacts with NPHP1. A, B: The association of RPGR with NPHP1 in bovine retina was analyzed by co-IP using appropriate antibodies. Precipitated proteins were analyzed by SDS–PAGE followed by immunoblotting with the indicated antibodies. C: Schematic representation of the primary structure of NPHP1. Abbreviations: NHD represents the nephrocystin homology domain; BD represents the binding domain; SH3 represents the Src homology domain. D: Interaction of GST-RPGR with ³⁵S NPHP1 was analyzed by GST pull-down assay. Amount of GST bound proteins was evaluated by Coomassie blue staining (asterisk; lower panel). E: This panel denotes expression of different domains of ³⁵S-NPHP1 (asterisks) used in the GST pull-down assay. Gels were analyzed by autoradiography. F: The interaction of different domains of NPHP1 with RPGR was analyzed by GST pull-down assay. Coomassie blue staining of the gels loaded with GST-RPGR protein was performed to evaluate the amount of protein in each experiment (lower panel). Molecular weight markers in kDa are shown on the left. Arrows indicate the specific protein bands representing the ³⁵S-NPHP1 deleted domains.