Akt1/protein kinase Bα is critical for ischemic and VEGF-mediated angiogenesis
J. Clin. Invest. Eric Ackah, et al. 115:2119 doi:10.1172/JCI24726 [Go to this article.]

Figure 5
Characterization of Akt-deficient ECs and fibroblasts. (A) Characterization of MLECs. Lung ECs were isolated from Akt1^–/–, Akt2^–/–, and WT littermates. Some Akt1^–/– ECs were reconstituted with a retrovirus expressing HA-tagged Akt1 (Akt1^–/–rcAkt1) or with GFP (Akt1^–/–rcGFP). WB, Western blot. (B) Characterization of mouse lung fibroblasts. (C) Relative expression of Akt isoforms in mouse cells. Lysates from MLECs, MLFs, and MASMCs were analyzed by SDS-PAGE and quantitative Western blot. Relative protein amounts of Akt1, Akt2, and Akt3 in the cells were quantified using standard curves obtained by running recombinant mouse Akt1, Akt2, and Akt3 proteins. (D) Basal phosphorylation of Akt and Akt substrates in MLECs. Cells were serum-starved for 48 hours and cell lysates subjected to SDS-PAGE and Western blot using the indicated antibodies. (E) Densitometric quantitation of p-protein to total protein for eNOS, GSK3β, and MDM2. For p-FKHR, data is expressed relative to β-actin loading control.