Figure 4.

The TOR pathway functions via JH to regulate Vg synthesis. (A) JHAMT expression in C. lividipennis females supplied with diets A1–A9 (as defined in Figure 1 legend). (B) JHAMT expression in females supplied with artificial diets that vary in AAs (as defined in Figure 2 legend). (C) JHAMT expression in females treated with rapamycin and ethanol (control). (D) JHAMT expression in C. lividipennis treated with dietary dsRheb, dsTOR, and dsS6K. (E) Vg expression in females treated with 0.2 nM rapamycin and rapamycin with methoprene (100 ng/nl); the ethanol + acetone treatment served as solvent control. Panels (F–H) show Vg expression in females fed on dietary dsRheb, dsTOR, and dsS6K and exposed to methoprene or acetone. Females treated with dietary dsGFP and acetone served as a control. Panels (I–L) show Vg protein levels in C. lividipennis (n = 15) treated with rapamycin, methoprene, dsRheb, dsTOR, and dsS6K. Proteins were detected with anti-Vg antibodies and visualized by western blot analysis; antiserum to β-actin was used as a loading control. Each treatment and control consisted of three independent biological replicates. Error bars represent means ± SEM. Columns labeled with different lowercase letters indicate significant differences at p < 0.05 by Student’s t-test.