Glucosylceramide synthase maintains influenza virus entry and infection
Fig 5
UGCG maintains optimal entry of VLPs bearing the glycoproteins of VSV, WSN influenza virus and EBOV.
VLPs were generated on an influenza virus βlaM1 backbone with the indicated viral glycoprotein. VLPs were added to prechilled cells which were then centrifuged at 4° for 1 hour. Next the cells were incubated for 3 hours at 37°, and then for 1 hour at room temperature in the presence of the βlaM substrate CCF2. Cells were washed, stored in the dark at room temperature, and (the following day) harvested, fixed, and analyzed for β-lactamase activity via flow cytometry. (A,B) Entry by VLPs bearing VSV G was reduced in HEK 293 UGCG KO cells, but unaffected in A549 KO cells. WSN influenza virus glycoprotein-mediated entry was reduced in both KO cell lines, consistent with the findings in Fig 4. Entry mediated by the EBOV glycoprotein was reduced in both 293 and A549 UGCG KO cells, and to a greater extent than seen with VLPs bearing the glycoproteins from WSN or VSV (mean ± SE; n = 6). ** p<0.01 using a Mann-Whitney non-parametric test.