Fig. 2

Yeast larvicide sema.460 effectively silences the sema1a target gene and induces severe neural defects in the Ae. aegypti L4 brain. Larval consumption of dried inactivated sema.460 tablets results in silencing of sema1a (b), which is normally expressed at high levels throughout the L4 larval brain ([24]; control-treated animal is shown in a for comparison). These results were quantified through analysis of mean gray value comparisons of sema.460- versus control-treated larvae (c; n = 53 sema.460-treated L4 brains; n = 57 control-treated L4 brains; data are represented as average mean gray values in c, as well as d3 and e3). Average mean grey value analyses (compiled from three biological replicate experiments with 65 control-treated larvae in d3 and n = 51 sema.460-treated larvae in e3) indicated that levels of nc82, which marks synaptic active zones (white in d1, e1; red in d2, e2), were significantly reduced in the synaptic neuropil of L4 larvae, while TO-PRO nuclear staining (blue in d2 and e2) levels were not significantly different. ***P < 0.001 with respect to control-fed larval brains (c, e3, analyzed by t-test). Representative L4 brains are oriented dorsal upward in this figure. Abbreviations: SOG, sub-esophageal ganglion; SuEG, supra-esophageal ganglion; LAL, larval antennal lobe; OF, olfactory foramen; OL, optic lobe