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. 2005 Sep 8;102(38):13502–13507. doi: 10.1073/pnas.0501902102

Fig. 3.

Fig. 3.

Angptl1 and Angptl2 bind to endothelial cells, and possess antiapoptotic activity through the PI3-K/Akt pathway. (A) Western blotting analysis with an anti-FLAG antibody with (Left) or without (Right) 2-mercaptoethanol (2ME). (B and C) Binding of COMP-Angptl1-FLAG and Angptl2-FLAG to HUVECs. The FITC intensity indicates cells bound by FLAG-tagged proteins. With both COMP-Angptl1 (Left) and Angptl2 (Right), intensities increased in a dose-dependent manner with saturation at ≈2 μg/ml. (D) Phosphorylation assay of ERK1/2 and Akt. HUVECs were treated with COMP-Angptl1 or -Angptl2 proteins. Immunoblotting was performed with anti-pERK1/2 or anti-pAkt antibody. Total amounts of ERK1/2 or Akt proteins were monitored by reprobing membranes with anti-ERK1/2 antibody and anti-Akt antibody. (E) TUNEL assay. HUVECs were cultured in the presence of either vehicle (PBS) or 0.5 μg/ml of COMP-Angptl1 or -Angptl2. In some experiments, cells were incubated with DMSO or 5 μg/ml PD980059 or LY294002. Data shown is the average of 6 fields each in six independent experiments (n = 36). *, P < 0.03 (compared with PBS in each group).