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. 2021 Jul 1;22(9):e52547. doi: 10.15252/embr.202152547

Figure 2. Type I IFN treatment solely at time of infection inhibits LY294002‐mediated reactivation of HSV‐1 in primary sympathetic SCG neurons.

Figure 2

  1. Schematic of the primary postnatal sympathetic neuron‐derived model of HSV‐1 latency.
  2. Reactivation from latency is quantified by Us11‐GFP expressing neurons following addition of the PI3K inhibitor LY294002 (20 μM) in the presence of WAY‐150168, which prevents cell‐to‐cell spread. The arrow indicates the time of LY294002 treatment at 5 days post‐establishment of latency. n = 9 biological replicates.
  3. Number of Us11‐GFP expressing neurons at 3 days post‐LY294002‐induced reactivation in P6 SCG neuronal cultures infected with HSV‐1 in the presence or absence of IFNα (600 IU/ml), then treated with an α‐ΙFNAR1 neutralizing antibody. n = 9 biological replicates.
  4. RT–qPCR for viral mRNA transcripts at 3 days post‐LY294002‐induced reactivation of SCGs infected with HSV‐1 in the presence or absence of IFNα. n = 9 biological replicates.
  5. RT–qPCR for viral mRNA transcripts at 20 h post‐LY294002‐induced reactivation in SCGs infected with HSV‐1 in the presence of absence of IFNα. n = 9 biological replicates.
  6. Relative amount of viral DNA at time of reactivation (8 dpi) in SCG neurons infected with HSV‐1 in the presence or absence of IFNα (600 IU/ml). n = 9 biological replicates.
  7. Quantification of vDNA foci detected by click chemistry at time of reactivation (8 dpi) in SCG neurons infected with HSV‐1 in the presence or absence of IFNα (600 IU/ml). n = 60 genomes from 3 biological replicates.
  8. LAT mRNA expression at time of reactivation (8 dpi) in neurons infected with HSV‐1 in the presence or absence of IFNα (600 IU/ml). n = 9 biological replicates.

Data information: Data represent the mean ± SEM. Statistical comparisons were made using a Mann–Whitney test (ns not significant, **P < 0.01, ****P < 0.0001).

Source data are available online for this figure.