Ly294002 inhibits FGF2-induced DNA biosynthesis.

(A) Serum-deprived cells were stimulated (black bar) or not (white bar) with FGF2 (10 ng/mL) in the presence (hatched bars) or absence of Ly294002 (10 μM). DNA biosynthesis was determined by measuring [³H]TdR incorporation during a 20-hour pulse as described in “Materials and methods.” Data are expressed as the means of triplicate values ± SEM and are representative of 6 experiments. (B) Detection of phosphorylated Akt. ACE cells were stimulated for 10 minutes with FGF2 (10 ng/mL) in the presence of various concentrations of Ly294002 (1 μM to 10 μM). Cell lysates were analyzed by Western blotting, using polyclonal antibodies recognizing phosphorylated or total Akt. Results are representative of 4 experiments. (C) Detection of phosphorylated and total Akt. Serum-deprived ACE cells were incubated with anti-FGF2 and anti–FGF-R1 antibodies or nonimmune IgG. Cell lysates were analyzed by Western blotting, using polyclonal antibodies against phosphorylated and total Akt. Results are representative of 3 experiments.