The effect of adenovirus-transduced cyclin A gene on DNA synthesis in Hcy-treated HUVECs.
HUVECs were infected with control adenovirus (Ad vector), adenovirus expressing cyclin A (Ad cyclin A) or adenovirus expressing β-gal (Ad β-gal) at the indicated MOI for 24 hours before being challenged with Hcy. Cells were then treated with or without 50 μM ofdl-Hcy in control medium for another 24 hours. (A) Western analysis; total protein (50 μg) from each sample was analyzed with antibodies against cyclin A. (B) Staining for β-gal; cells were infected with Ad β-gal at the indicated MOI and stained for β-gal. Photograph was taken with × 100 magnification. (C) Thymidine uptake; cells treated for 24 hours were metabolically labeled with 1 μCi (0.037 MBq)/mL [3H]-thymidine during the last 3 hours. [3H]-thymidine incorporation was measured in a liquid scintillation counter. ■, control; ▪, Hcy, 24 h. Values represent mean ± SD from 3 separate experiments from 3 wells (n = 9).
