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. 2020 May 27;16(5):e1008539. doi: 10.1371/journal.ppat.1008539

Fig 5. The NAD salvage pathway controls metabolism, and survival of adult and immature S. japonicum.

Fig 5

(A) Intracellular NAD levels in female S. japonicum cultured for 48h with vehicle or 250nM FK866 ± 2mM NMN or Trp. n = 5–6 wells/group with 1 female parasite/well. (B) Egg production by female S. japonicum cultured for 48h with vehicle or 250nM FK866 ± 2mM NMN or Trp. n = 4–5 wells/group with 1 female parasite/well. (C-E) Loss of membrane integrity in day 7 FK866-exposed male S. japonicum. Representative H&E stained cross sections (C) and SEM images (E) of male S. japonicum cultured for 7 days with vehicle or 250nM FK866. Severity of tissue damage reported as vacuolization scores (D) as determined by blinded assessment of the H&E cross sections (see S2 Fig for pathology scoring methodology and S3 Fig for additional SEM micrographs). n = 7–10 worms/group/analysis. (F) Parasite mobility by adult S. japonicum cultured for 0–12 days with vehicle or 250nM FK866 ± 2mM NMN or Trp. n = pooled averages from 3 individual experiments consisting of 3 wells/condition with each well containing 2 male and 2 female parasites. (G) Survival of S. japonicum cultured for 12 days with vehicle or 250nM FK866 ± 2mM NMN or Trp. n = 12 parasites (2 males + 2 females/well in triplicate wells). (H) Parasite mobility by S. japonicum schistosomula cultured for 0–10 days with vehicle or 250nM FK866 ± 2mM NMN or Trp. n = 3 wells/group with 6–9 schistosomula/well. (I) Survival of S. japonicum schistosomula cultured for 10 days with vehicle or 250nM FK866 ± 2mM NMN or Trp. Data are shown as the percentage of parasites alive at each timepoint. n = 6–9 parasites/group. Data are representative of 2 (A-E, H-I) or 3 (F-G) independent experiments. Data shown as the mean ± SD (A-B, D, H) or mean ± SEM (F) with individual samples shown in squares. Statistical tests were performed using one-way ANOVA multiple comparison tests for experiments with more than two groups, two-tailed Student’s t-test for experiments with two groups or a Log-rank (Mantel-Cox) test for the survival experiments.