Invasion of midgut epithelial cells by a persistently transmitted virus is mediated by sugar transporter 6 in its insect vector
Fig 1
LsST6 interacted with viral proteins in vivo and in vitro.
(A) Interaction between LsST6 and the outer capsid or nucleocapsid protein of different viruses detected using an in vivo yeast two-hybrid assay. Yeast strain NMY51 was cotransformed with LsST6 and the respective viral proteins. After 10−1 to 10−4 times dilution, the yeast cells were plated onto DDO (SD-trp-leu) and QDO (SD-trp-leu-his-ade-20 mM 3-AT) medium plates, respectively. Clones grown on DDO were selected for β-galactosidase activity assay. Large T + P53 was used as the positive control; Large T + LsST6 served as the negative control. (B) Confirmation of the interaction by in vitro coimmunoprecipitation (Co-IP). Sf9 cells were cotransfected with the respective recombinant bacmids (LsST6 fused with His-tag and viral proteins linked with Myc-tag) for protein expression. After 72 h, cells were harvested, lysed in lysis buffer, then incubated with anti-Myc antibody and protein A/G plus agarose beads for IP. The anti-Myc antibody or anti-His antibody was used for western blots.