Fig 2: Schematic diagram of the absence of SPP in peripheral sensory neurons of Avil-SPP-/- mice and its effect on latency-reactivation, not primary ocular infection.SPP is present in the eye of Avil-SPP-/- mice and virus replication is not affected. As shown, HSV-1 replicates normally in peripheral tissue due to the presence of SPP and moves to peripheral neurons via retrograde transport. However, knockdown of SPP results in inhibition of HSV-1 replication in Avil-SPP-/- neurons and blocking/reduction of latency establishment, thus affecting transport of virus to site of infection and reactivation.

Fig 3: Expression of CD8a and PD-1 transcripts in TG of Avil-SPP-/- infected mice during primary infection.Isolated RNA from total TG and neuronal fraction (see Fig 3) was used to measure CD8a and PD-1 transcript expression by qRT-PCR. The ratio of expression for each mRNA transcript was normalized to its expression in its respective uninfected group. GAPDH expression was used to normalize relative expression of each transcript. Neuronal RNA was isolated from four combined TG, while total TG RNA was isolated from individual TG. Each bar represents the mean ± SEM from 12 mouse TG for neurons and 12 TG from six mice for total TG RNA.

Fig 4: Effect of SPP deficiency on virus reactivation in infected mice.Avil-SPP-/- and control mice were ocularly infected with 2 X 105 PFU/eye of HSV-1 strain McKrae. TG from infected mice were harvested on day 28 PI and the explant reactivation assay was performed. Data points indicate the day of virus reactivation. Each point is the mean ± SEM from 34 TG.

Fig 5: Expression of gB, gK, and ICP0 transcripts in corneas and TG of infected mice.Pax6-SPP-/- (gray bars) and WT control mice (open bars) were ocularly infected with 2 X 105 pfu/eye of HSV strain McKrae. Corneas and TG were harvested on day 4 PI. RNA from corneas (panel A) or TG (panel B) were isolated as described in Materials and Methods. gB, gK, and ICP0 transcripts were measured by qRT-PCR and copy numbers for each transcript were calculated using a standard curve generated from their respective plasmid. Each bar represents the mean ± SEM from 10 corneas or 10 TG.