Skip to main content
. 2020 Mar 25;12(4):359. doi: 10.3390/v12040359

Figure 3.

Figure 3

Lack of apoptosis in stem cell-derived human neural cultures infected with lyssavirus strains. HDF51i-509-NPCs and H9-NPCs were differentiated for 24 days and 29 days, respectively. These neural cultures were infected with different lyssavirus strains for 72 h at virus:cell ratio of 1 and examined for apoptosis by TUNEL staining (TMR-red). (a) Representatives images of HDF51i-509-NPC-derived neurons identified by anti-neurofilament immunofluorescence (NF, green) stained with TUNEL (red) and DAPI (blue) for nuclei. Infection with each lyssavirus strain was identified by anti-nucleoprotein staining (magenta). Images were taken with 20× objective and are maximum intensity projections of Z-stacks. Scale bar 50 µm. Quantification of apoptotic DNA fragmentation using TUNEL staining in (b) H9-NPC-derived neurons and (c) HDF51i-509-NPC-derived neurons. At least 500 neurons were quantified per sample, n = 3. No statistical significance in apoptotic cell death observed between uninfected and lyssavirus infected neurons. qPCR analysis of apoptotic (caspase9) and necrosis (RIP1) genes in total RNA from (d) H9-NPC-derived and (e) HDF51i-509-NPC-derived neural cell cultures. No statistical significance difference is observed in the expression of caspase9 and RIP1 genes in uninfected and lyssavirus infected neural cultures.