Activated Akt-1 does not protect against NaSal-induced apoptosis. (A) Effect of NaSal on Akt-1 expression and activation. Monocyte-differentiated macrophages were treated with NaSal (20 mM) for 12 to 48 hours, then cell lysates were collected and Western blot analysis performed using the antibodies to phosphorylated and total Akt-1. No indicates no NaSal added. (B-D) Seven-day differentiated macrophages were untreated (Normal) or infected with adenoviral vectors (100 moi) expressing the control β-galactosidase, Bcl-x_L, or activated Akt-1 (MyrAkt-1) for 24 hours. Where indicated, NaSal (20 mM) was added for an additional 48 hours. The cells were harvested and analyzed for Δψ_m (mean fluorescence), cell death (PI-positive cells), or apoptosis (DNA fragmentation). Values represent the mean ± 1 SE and were compared for statistical significance by Student t test for paired samples. ** indicates P < .01 compared with control Adβgal. Shown is a representative of 2 experiments performed in triplicate.