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. 2020 Apr 15;39(10):e103758. doi: 10.15252/embj.2019103758

Figure EV1. IFNγ‐mediated susceptibility of the type II Pru strain is neither dependent on MYR1‐dependent parasite secreted factors nor on L‐Trp breakdown by primary human fibroblasts.

Figure EV1

HFFs were pre‐stimulated for 24 h with 10 U/ml IFNγ followed by infection with the indicated strains.
  • A
    Representative images of the plaque assay described for Fig 1A. Left panel is for unstimulated HFFs infected with either RH (top) or Pru (bottom), and right panel is for IFNγ‐stimulated HFFs infected with RH (top) and Pru (bottom). Scale bar is 100 μm.
  • B
    Plaque assays were performed with PruΔku80Δhpt (Pru) and PruΔku80ΔhptΔmyr1 (PruΔmyr1) strains, and plaque numbers were counted and plaque areas were measured after 6 days. Plaque numbers and plaque areas are averages from eight and six biological replicates, respectively.
  • C
    Representative images of the plaque assay described for Fig 1E. Left panel is for unstimulated HFFs infected with Pru WT (top), Pru Δgra15 (middle), or Pru Δgra15 + GRA15 (bottom) and right panel is for IFNγ‐stimulated HFFs infected with Pru WT (top), Pru Δgra15 (middle), or Pru Δgra15 + GRA15 (bottom). Scale bar is 100 μm.
  • D
    Representative images of the plaque assay described for Fig 1F. Left panel is for unstimulated HFFs infected with RH (top) or RH + GRA15II (bottom), and right panel is for IFNγ‐stimulated HFFs infected with RH (top) and RH + GRA15II (bottom). Scale bar is 100 μm.
  • E
    Cells were supplemented with media containing 0.6 mM L‐Trp, stimulated with IFNγ for 24 h, and infected with RH or Pru parasites. Plaque number was measured 4 (RH) or 6 days (Pru) p.i. Experiments were performed two times.
  • F
    L‐Kynurenine was measured from HFF culture supernatant as a marker of IDO activity. Experiment was done two times.
  • G
    Number of total host nuclei counted per experiment for Fig 1C and H for each indicated strains and conditions. Experiments were performed six times with RH and Pru WT and three times with RH + GRA15II and Pru Δgra15
  • H
    Measurement of infectivity of Pru WT and Pru Δgra15 by plaque assay in unstimulated HFFs using equal number of parasites (250).
Data information: Data are represented as mean ± SEM.Source data are available online for this figure.