Figure 3
From: CdGAP maintains podocyte function and modulates focal adhesions in a Src kinase-dependent manner
β-PIX interacts with the basic region of CdGAP. (A) HEK293 cells overexpressing myc-CdGAP and GFP-β-PIX were lysed and cell lysates were immunoprecipitated with anti-GFP antibody (for β-PIX) followed by immunoblotting for myc (for CdGAP). A representative immunoblot is shown. (B) Densitometric quantification of IP: normalized to total expression of myc-CdGAP in (A). (C) Schematic representation of the CdGAP deletion mutants, which were used in the co-immunoprecipitation experiments. (D) Cell lysates of HEK293 cells overexpressing GFP-CdGAP deletion proteins and full-length mCherry-β-PIX were immunoprecipitated with anti-GFP antibody (for CdGAP) followed by immunoblotting for mCherry (for β-PIX). (E) Densitometric quantification of IP: normalized to total mCherry-β-PIX expression in (D). (F) Cell lysates of HEK293 cells overexpressing myc-CdGAP deletion proteins and full-length GFP-β-PIX were immunoprecipitated with anti-GFP antibody (for β-PIX) followed by immunoblotting for myc (for CdGAP). (G) Densitometric quantification of IP: normalized to total myc-CdGAP expression in (F). n = 5 (B); 2–4 (E), 2–3 (C). Statistically significant differences (****P < 0.0001), assessed by the Student’s t-test test (B), are indicated. EV: Empty vector; BR: basic-rich; PRD: proline-rich domain; C-term: C-terminus.
