Figure 3.

Productive EBOV infection of iPSC-HLCs

(A) RNA-FISH analysis of BU2 EBOV-infected iPSC-HLCs at the indicated time points using probes directed against EBOV VP35 mRNA. Scale bar represents 20 μm. Images are representative of BU1, BU2, and BU3 samples (n = 1 technical replicate per iPSC donor per time point for a total of three independent replicates per time point).

(B–F) Transmission electron microscopy of BU2 EBOV-infected iPSC-HLCs at 1 and 2 dpi. Cells were infected with an MOI of 3. Image is representative of infections in BU1, BU2, and BU3 (n = 1 replicates per iPSC donor per time point for a total of three independent replicates per time point). (B) Overview of BU2 EBOV-infected cell at 2 dpi. Circled area indicates the accumulation of filamentous viral nucleocapsids into inclusions. (C) Higher magnification of cross-sectioned (left) and longitudinal sectioned (right) viral inclusions. Single nucleocapsids are marked with yellow and red asterisks, respectively, to visualize the hexagonal pattern. (D and E) Release of viral particles at 1 dpi (D) and 2 dpi (E). Circled area in (E) indicates viral inclusions. (F) Arrows indicate mature, budding Ebola virions. Cells were fixed at 2 dpi. Images were taken from cells derived from different donors: (A–D) donor BU2, (E) donor BU1, (F) donor BU3.