Skip to main content
. 2011 Jan 31;108(7):2957–2962. doi: 10.1073/pnas.1009395108

Fig. 5.

Fig. 5.

Shp2 binding site in Gab1 is critical for VEGF-induced eNOS and PKA substrate phosphorylation. The ECs from the control and EGKO mice, or EGKO ECs overexpressing WT Gab1 or mutants and HUVECs overexpressing vector, Gab1 WT or mutants were serum-starved and stimulated with VEGF (50 ng/mL) for indicated time periods, followed by immunoblot analysis (A–C) and tube formation assay (D). (A) Immunoblot analysis of phosphorylations of eNOS and PKA substrate, and of Gab1 in control cells, parental and Gab1 WT, or indicated mutant-rescued EGKO cells. The differences in P-eNOS levels of Gab1 mutant-rescued EGKO cells were significant compared with Gab1 WT-rescued EGKO cells (P < 0.01). (B) Immunoblot analysis of anti-Gab1 or anti-PKA Cα immunoprecipitates from the lysates of HUVECs overexpressing Gab1. P85 was used as loading control. (C) Immunoblot analysis of anti-Gab1 immunoprecipitates from the lysates of HUVECs overexpressing vector, Gab1 WT, or indicated mutants. (D) Tube formation analysis of HUVECs overexpressing vector, Gab1 WT, or indicated mutants. Asterisks indicate statistical significance (*P < 0.05, **P < 0.01) for EGKO versus control.