Fig. 2 | Nature Communications

Fig. 2

From: Leishmania RNA virus exacerbates Leishmaniasis by subverting innate immunity via TLR3-mediated NLRP3 inflammasome inhibition

Fig. 2

LRV increases L.g. virulence in vitro and in vivo. af C57BL/6 mice (n = 4 per group) were injected intradermally in the left ear with SP (ac) or metacyclic promastigotes (df) of L.g.− or L.g.+ and ear thicknesses were followed weekly (a, d). At 4 w.p.i., parasite titers were determined in the infected ear (b, e) and draining lymph node (c, f). gm WT BMDMs were infected with metacyclic promastigotes of L.g.− and L.g.+ at a MOI of 1. Cells were washed after 1 h of infection, and kept in culture for 1, 24, 48, 72, or 96 h. g, h In vitro killing assay (Panotico Giemsa) showing the percentage of infected cells (g) and the average number of parasites per cell (h) in the end of each time point are shown. (im) FACS analysis of BMDMs infected with L.g.− and L.g.+ that were pre-incubated with CFSE dye prior to infection. The representative contour plot i and the percentages of alive and dead infected (j, k) or non-infected (l, m) cells are shown. One representative of three (af) or two (gm) independent experiments performed is shown, with biological (af) or technical (gm) replicates. The results are shown as mean ± SD. Statistical analysis was performed by two-way ANOVA with Bonferroni’s multiple comparison test. P < 0.05 was considered statistically significant (*)

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