The ovariole infection was in sync with BmVg expression.

(A) BmVg protein content in the ovarioles at different stage was determined by western blot using polyclonal antibody against BmVg (anti-BmVg) with B. mori α-tubulin (Bm-α-tubulin) as an internal reference. (B) Fold changes of BmVg content relative to Bm-α-tubulin in ovarioles. P1d, 1st day of pupation, P3d, 3rd day of pupation, P5d, 5th day of pupation, P7d, 7th day of pupation. (C) Fold changes of pathogen load indicated by the copy numbers of N. bombycis β-tubulin in ovarioles at different stages. Bars represent the average fold change per experiment ± SD. (D) IFA analysis of BmVg localization on the surface of N. bombycis spores. (D1) N. bombycis spores isolated from female pupae were labeled with the anti-BmVg. (D2) N. bombycis spores isolated from female pupae were incubated with negative serum as a control. (D3) N. bombycis spores isolated from male pupae were incubated with the anti-BmVg. (D4) N. bombycis spores isolated from male pupa were incubated with hemolymph of female pupa and labeled with the anti-BmVg. Nuclei were stained using DAPI (blue). The green fluorescence indicates the BmVg labeled by anti-BmVg conjugated with Alexa Fluor 488. Bars, 5 μm.