Effect of JH treatment on the intracellular levels of calcium, cAMP, DAG, and IP3 in mosquito Aag2 cells. Cells were treated with the indicated chemicals (1 µM) or ethanol (0.1%) as a solvent control. (A) Cytoplasmic calcium concentration was measured using the Fluo-8 AM fluorescent calcium indicator and a microplate reader. Values are expressed as relative fluorescence intensity. (B) Intracellular cAMP contents were assessed after the JH treatment. The adenylate cyclase activator forskolin was used as a positive control. (C and D) The amounts of DAG (C) and IP3 (D) were determined in cells 5, 10, 30, 45, and 60 min after the addition of JH-III. All experiments were repeated three times with similar results. Data are reported as the mean and SD of triplicate samples from a representative experiment.