Figure 6. TEP1 localizes to hyphae and hyphal bodies.

(A) LacZ (control) and (B) TEP1 kd mosquitoes were injected each with 200 conidia of GFP-expressing B. bassiana. Abdomens were dissected 48 h later and immunostained with TEP1 antibody. Shown are bright field (Bf) and merged bright field and red channel (Bf+red) confocal sections and Z projections (Z-proj) of whole stacks. (A) In control mosquitoes TEP1 staining was observed on established hyphae (Aii, filled arrowheads), young branching hyphae (Aii and Aiii, open arrowheads) and hyphal bodies (Aiii, inset and filled arrowhead). Most TEP1-stained hyphae did not exhibit melanin formation at that time point (compare Ai and Aii), except in rare cases where TEP1 signal overlapped with a thin layer of melanin (Ai and Aii, arrows with open heads). TEP1 was not detected on heavily melanized hyphal surfaces probably because it was masked by melanin (Ai and Aii, arrows with filled heads). (B) TEP1 staining of hyphae was completely abolished in TEP1 kd mosquitoes (compare Aii with Bii, and Aiii with Biii). Consequently, melanin did not form over these hyphal surfaces but was present only on early fungal stages including germinating spores and germ tubes (Bi, open arrowheads). Note the abundance of hyphal bodies in TEP1 kd (Biii, arrows) compared to controls. TEP1 (red) GFP-expressing B. bassiana (green), nuclei (blue). All scale bars are 50 µm.