Figure 2.

PDIA3 inhibits EBOV entry. (a) HIV-1 firefly luciferase reporter viruses pseudotyped with GP or GP∆MLD were produced from HEK293T WT, PDIA3-overexpressing, or PDIA3-KO cells. After infecting HEK293T cells with an equal number of these different viruses, viral entry was determined by measuring intracellular luciferase activities. Viral entry is shown as relative values, with the entry of GP∆MLD-pseudotyped viruses in the absence of PDIA3 normalized to 100%. (b) EBOV virus-like particles (VLPs) were produced from HEK293T WT, PDIA3-overexpressing, or PDIA3-KO cells after expression of GP and EBOV-VP40. VLPs were purified by ultra-centrifugation and analyzed by WB. Levels of GP in virions were further quantified from these Western blots and are presented as relative values as previously. (c) EBOV replication and transcription-competent virus-like particles (trVLPs) were produced and passaged two times (p0, p1, p2) in HEK293T cells in the presence or absence of PDIA3. EBOV replication was determined by measuring intracellular Renilla luciferase activity. Viral replication was also measured in the absence of EBOV-L, which is a negative control. Results from three independent experiments are presented. Error bars in A and B represent SEMs calculated from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns (p > 0.05).