Involvement of Foxo transcription factors in angiogenesis and postnatal neovascularization
J. Clin. Invest. Michael Potente, et al. 115:2382 doi:10.1172/JCI23126 [
Go to this article.]
Figure 1Foxo1 and Foxo3a are the predominant Foxo transcription factors in endothelial cells. (
A) Statistical summary of the Foxo expression profile as assessed in a microarray analysis of total RNA isolated from HUVECs. Data are presented as mean ± SEM;
n = 4. The values on the
y axis represent a ratio normalized to the mean fluorescent intensity of all genes on the chip. (
B) Cell fractions were prepared from HUVECs that were either left untreated, serum starved for 24 hours, or transfected with Flag-Foxo1, HA-Foxo3a, or HA-Foxo4 and subjected to SDS-PAGE. Western blot analysis was performed using the indicated antibodies. Tagged, epitope-tagged; endo., endogenous. (
C) HUVECs were serum-starved for 24 hours and then either left untreated or stimulated with VEGF (50 ng/ml) for the indicated times. Cell lysates were subjected to Western blot analysis with antibodies against phospho-Foxo1 (Thr 24), phospho-Foxo3a (Thr 32), or total Foxo1. (
D) HUVECs were serum starved for 24 hours, then incubated with 10 μM LY294002 for 1 hour and stimulated with VEGF (50 ng/ml) for the indicated times. Cell lysates were analyzed by Western blot analysis using an antibody directed against phospho-Foxo1. The total level of protein was assayed by Western blot analysis using an anti-Foxo1 and anti-Akt antibody. FCS (20%) was used as a positive control.
