FIGURE 2.

D-NAAM mRNA expression is regulated by oxidative stress. A, Drosophila S2 cells were cultured under the indicated growth conditions, and the expression of D-NAAM mRNA was analyzed using real time PCR. The data are presented as fold change in D-NAAM expression after standardizing to ribosomal protein L32. The treatments were as follows: C, control, complete SFM media; DS, DS2 media (media lacking protein factor additives); SFM 1:4, SFM medium diluted 1:4 with phosphate-buffered saline (nutrient deficient media), 18 h; DS 1:4, DS medium diluted 1:4 with phosphate-buffered saline, 18 h; FCS, 10% fetal calf serum, 4 h; LY, 10 μm LY294002, 18 h (PI3K inhibitor); HS1, heat shock, 2 h at 37 °C and 24 h recovery; HS2, heat shock, 2 h at 37 °C; H₂O₂, 15 μm, 18 h; Sor, 0.5 m sorbitol, 18 h; Ani, 10 μg/ml anisomycin, 18 h; DMSO, 0.1% Me₂SO, 18 h (control); Rapa, rapamycin, 0.2 or 0.5 μg/ml, 18 h; Res, 100 μm resveratrol, 18 h; NAM, 40 mm nicotinamide, 18 h; Sir, 50 μm sirtinol, 18 h. B, Drosophila S2 cells growing in complete SFM medium were treated as indicated or cells were irradiated and left to recover for 24 h. The cells were analyzed for changes in D-NAAM mRNA expression as in A. C, Drosophila S2 cells were treated with the indicated concentrations of H₂O₂ for the indicated times, and the D-NAAM mRNA levels were analyzed as in A.