Figure 2.

Knockdown of Aacec B in Ae. aegypti pupae leads to high pupal mortality and the emergence of deformed adult. (a) RT-qPCR analysis of Ae. aegypti cecropins expression in uninjected control, GFP dsRNA-injected control and Aacec B dsRNA-injected pupae. P_< 0.5: Pupae were injected within 0.5 hr after larval-pupal ecdysis; P₁₂–P₄₈: 12–48 hrs after pupation. The relative expression levels are expressed as means ± SD (n = 3), with uninjected pupae at <0.5 hr after injection as the calibrator. Asterisks indicate significant differences (*p < 0.005; **p < 0.001) from the relative mRNA levels of the uninjected control at each time-point. (b) The cumulative mortality rates (%) of uninjected control (blue line), GFP dsRNA-injected control (red line), Aacec B dsRNA-injected (green line), and Aacec B dsRNA +Aacec B peptide-injected (orange line) Ae. aegypti pupae at various times after injection. P_< 0.5: Pupae were injected within 0.5 hr after larval-pupal ecdysis; 24–120: 24–120 hrs after larval-pupal ecdysis. Each group consisted of 30 pupae in one experiment and 5 replicates were conducted. The range bars indicates the standard deviations of the means. (c) An adult mosquito from the Aacec B dsRNA-injected pupae that was unable to detach from the exuvia. (d) An adult mosquito with deformed body parts. Arrows (↑) indicate curved wings and arrow heads (▲) indicate curved legs. (e) A normal mosquito that emerged from the GFP dsRNA-injected pupae.