Altered Immunity in Crowded Locust Reduced Fungal (Metarhizium anisopliae) Pathogenesis
Figure 5
Locust GNBP3 affected attacin expression and was susceptible to humoral protease.
(A) Immunofluorescence observed the locust GNBPs binding to fungi cell wall by confocal microscope. DIC: differential interference contrast; FITC: fluoresceinisothiocyanate staining; DAPI: 4′,6-diamidino-2-phenylindole staining; Ctr: Drosophila hemolymph injected with M. anisopliae was used as negative control for examining reactivity of anti-GNBP1 and anti-GNBP3 pAbs. White arrow indicated positive signals. (B) Locust attacin expression in fat body after RNAi knockdown of GNBP1 or GNBP3 was examined by quantitative real time PCR. Fold change was normalized to GFP knockdown locusts. (Mann-Whitney U test, nGNBP1 = 9, P<0.001; nGNBP3 = 9, P<0.001) (C) After 2-hours incubation of laminarin-stimulated hemolymph at room temperature, the proteolysis samples of GNBP1 and GNBP3 proteins in supernatant and clot were analyzed by immunoblot. Ctr: before incubation; In: 2-hours incubation; Pe: pellets after incubation. The black arrow indicates the degraded fragment of GNBP3 recognized by anti-N-terminal pAbs.