Fig. 3: Direct association of RdFV CP or RGDV P8 with HongrES1 on sperms in infected testes of male R. dorsalis.

Association of RdFV CP (a) or RGDV P8 (b) with HongrES1, as determined by immunofluorecense microscopy. The dissected testes from 30 RdFV- or RGDV-positive males were immunostained with HongrES1-FITC (green) and CP-rhodamine (red) (a), or with HongrES1-FITC (green) and P8-rhodamine (red) (b). Ps, punctate structure. Fs, filamentous structure. Bars, 10 μm. c The mean percentages of CP- or P8-formed structures colocalized with HongrES1 in respective infected testes (n = 30). Means (±SD) represent three biological replicates (two-tailed t-test). Ns, not significant. Association of RdFV CP or RGDV P8 with HongeES1 on the dissected sperms, as determined by immunofluorecense microscopy. The dissected sperms from 30 RdFV- or RGDV-positive testes were stained with HongrES1-FITC (green), CP-rhodamine (red) and nuclear dye DAPI (blue) to indicate sperm head (d), or with HongrES1-FITC (green), P8-rhodamine (red) and DAPI (blue) (e). Sh, sperm head. Bars, 10 μm. f The mean percentages of sperms (n = 100) associated with capsid proteins and HongrES1. Means (±SD) represent three biological replicates (two-tailed t-test). Ns, not significant. g Electron micrograph showing the sperms in virus-free testes. Bar, 500 nm. Electron micrograph showing the association of RGDV virions (h) or virus-containing tubules (i) with the plasma membrane of sperms in RGDV-infected testes. Bars, 500 nm. j–m Immunoelectron microscopy showing the association of HongrES1 with RGDV particles and sperms. Male testes were immunolabeled with HongrES1-specific IgG as the primary antibody, followed by treatment with 15-nm gold particle-conjugated IgG as the secondary antibody. Red arrows indicate gold particles. Vi, virions; Tu, tubule. Bars, 100 nm.