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. Author manuscript; available in PMC: 2009 Jan 1.
Published in final edited form as: J Insect Physiol. 2007 Oct 2;54(1):231–239. doi: 10.1016/j.jinsphys.2007.09.007

Fig. 3.

Fig. 3

JHIII treatment changes the expression profiles of the TOR signaling pathway components in fat bodies of small mosquitoes. Juvenile hormone III (JHIII) was topically applied to newly eclosed small mosquitoes, and fat bodies were subsequently dissected at different time points before and after a blood meal. (A) AaiCAT2 gene expression was analyzed using AaiCAT2-specific real-time PCR primers for standard mosquitoes (solid line), small mosquitoes (dotted-dashed line), and JHIII-treated small mosquitoes (dashed line). Values are means ± SD of triplicate samples from different cohorts. (B) AaiCAT2 protein expression in the fat body of female mosquitoes was assessed by Western blotting at different time points during the first gonotrophic cycle. The blot shown is representative of three independent experiments. (C) mRNA expression profiles of TOR in standard (solid line), small (dotted-dashed line) and JHIII-treated small (dashed line) mosquitoes were examined using real-time PCR. Values are means ± SD of triplicate samples from different cohorts. (D) The native TOR protein was examined by Western blotting using an anti-TOR polyclonal antibody preparation. An antibody against β-actin was used as a loading control. The blot shown is representative of three independent experiments. (E) Western blot analysis was used to assess the phosphorylation state of S6K (phospo-S6K) with a polyclonal antibody preparation against Thr(P)-388 of S6K (S6K-P) along with an antibody preparation against S6K (S6K) as the loading control. The blot shown is representative of three independent experiments. (F) Vg gene expression was analyzed using Vg-specific real-time PCR primers in standard mosquitoes (solid line), small mosquitoes (dotted-dashed line), and JHIII-treated small mosquitoes (dashed line). Values are means ± SD of triplicate samples from different cohorts. (G) Vg protein content in the fat body of female mosquitoes was determined by Western blotting at different time points during the first gonotrophic cycle. The blot shown is representative of three independent experiments.