Figure 2.

FHV-B2 over-expression in midguts of transgenic mosquitoes of line PUbB2 P61. (A) Western blot analysis of PUbB2 P61 females to detect FHV-B2 expression. Total protein extracted from five midguts or carcasses of sugarfed PubB2 P61 and HWE (negative control) mosquitoes was electrophoretically separated in 10% SDS-PAGE. B2 over-expression was detected with an IgG antibody specific for the v5 epitope tag. Total protein of SINV-TE3’2J-B2 infected cell culture medium was used as positive control. Lane 1: HWE midguts, 5 days post-eclosion; lane 2: PUbB2 P61 midguts, 5 days post-eclosion; lane 3: PUbB2 P61 midguts, 7 days post-eclosion; lane 4: SINV-TE3’2J-B2 culture supernatant, 36 h post-infection; lane 5: molecular size marker; lane 6: PUbB2 P61 carcasses, 1 day post-eclosion; lane 7: PUbB2 P61 carcasses, 5 days post-eclosion; lane 8: PUbB2 P61 carcasses, 7 days post-eclosion. (B) Detection of FHV-B2 protein in midguts by Immunofluorescence assay using anti-v5 IgG as detection antibody. (1) sugarfed HWE (negative control); (2) sugarfed PUbB2 P61 female; (3) PUbB2 P61 female at 5 days pbm; (4) DENV2-QR94 infected female at 5 days pbm; (5) close-up image (400×) of midgut epithelium over-expressing FHV-B2 in a sugarfed PUbB2 P61 female at 7 days post-eclosion; (6) close-up image (400×) of midgut epithelium of a sugarfed HWE female at 7 days post-eclosion.