L690 330

Cells were stimulated with PD184352 (Sigma-Aldrich), prostratin (Sigma-Aldrich), ingenol,3,20-dibenzoate (Enzo LifeSciences), L690,330 (Tocris), fluticasone propionate (Sigma), or Go6983 (Tocris) dissolved in dimethylsulfoxide (DMSO) or water (for L690,330). An equal amount of the vehicle (1:1,000) was used as solvent control.

Lithium chloride (LiCl; Sigma-Aldrich, St. Louis, MO) was dissolved in water to generate 1-, 0.5-, and 0.25-m stock solutions, which were added to the standard fly food in a 1:10 dilution to produce the final concentrations used in these studies. Newly eclosed flies were transferred to vials with LiCl-containing food and aged for 1–5 d before experiments were performed. For the experiments to evaluate the effects of inhibitors for GSK-3 or IMP, larvae were raised in the standard food containing 20 μm AR-A014418 (Sigma-Aldrich) or 0.5 mm L-690330 (Tocris, Bristol, UK) at 25°C. AR-A014418 and L-690330 were initially dissolved in DMSO and water to generate 20- and 0.5-m stock solutions, respectively. The eclosed adult flies were scored for their morphological phenotype and continued to be grown on the drug-containing food for 5–6 d before being assessed for locomotor activity.