Figure 7. Inhibition of Ae.aegypti midgut trypsin activity by Tsp_PR-secreted molecule(s).

(A) (left) Ovary development based on an arbitrary score of the ovary size at 6 days after a blood meal, 0 for small round follicles, 1 for intermediate size follicles, and 2 for fully developed follicles, with the elongated shape of normal mature eggs. Control, N = 34; Tsp_PR, N = 34, line represents the median, of three independent experiments. (right) Light microscopy picture of (a) a completely developed ovary follicle, which represents a score 2 (b) small round follicles, represent score 0. (B) Change in mosquito body weight after 1 hr (Control, N = 66; Tsp_PR, N = 74), (p=0.784) and 48 hr (Control, N = 58; Tsp_PR, N = 74) (p=0.001) of a non-infected blood meal (C) Trypsin in vivo enzymatic activity in midguts of mosquitoes treated or mock-treated with Tsp_PR secretome. Error bars represent ± SEM of three independent experiments. (D) Trypsin in vitro enzymatic assays of Tsp_PR’s ability to inhibit commercial trypsin activity. The activity was measured at various concentrations of trypsin. Tsp_PR represents the control group in which the fungus filtrate was added but no trypsin, and the absence of trypsin activity was experimentally confirmed (not shown). Error bars represent ± SEM of three independent experiments. *p<0.05, **p<0.01, ***p<0.01.