Fig. 2.

Invasion and transmission of H129-G4 in vitro (a–b) Invasion of H129-G4. Freshly isolated fetal mouse hippocampal and cortical neurons were seeded into one chamber of the microfluidic plate, and the first 24 h culture was termed as Day 1. H129-G4 was added at Day 8 into either the soma (blue) or axon terminal chamber (red) to a final concentration of 1 × 10⁷ pfu/ml (a). Images of GFP signal (upper panel) and phase contrast (lower panel) were obtained at 24 h post infection (hpi). The represent results from 3 microchannel plates are shown (b). The dotted lines indicate the borders between chambers and the microchannels. Scale bar = 100 μm (c–e) Transmission direction of H129-G4. Neurons were sequentially plated into the two chambers at Day 1 and Day 5, then H129-G4 was added at Day 12 into either the efferent (blue) or afferent chamber (red) to a final concentration of 1 × 10⁷pfu/ml (c). Images of GFP signal (upper panel) and phase contrast (lower panel) were obtained at 24hpi. The representative results from 3 microfluidic plates are shown (d), and the quantitative analysis were performed by counting the GFP labeled cell mounts in different chambers (e). The dotted lines indicate the borders between chambers and the microchannel. Scale bar = 100 μm