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. 2018 Sep 19;9:2073. doi: 10.3389/fimmu.2018.02073

Figure 4.

Figure 4

IDO1 but not ATG16L1 extensively participate in anti-T. gondii response in human cell lines. (A) WT or ATG16L1-KO HFFs, Huh7 or Hela cells were untreated or treated with IFN-γ for 24 h, and then lysates were detected by Western blot. (B) WT, IDO1-KO, or ATG16L1-KO HFFs, Huh7 or Hela cells were untreated or treated with IFN-γ for 24 h, and then infected with T. gondii. The parasite survival rate after 24 h post infection was measured by luciferase assay. (C) Fluorescence confocal microscopy of WT, ATG16L1-KO and IDO1-KO HFF, Huh7 or HeLa cells stimulated by IFN-γ for 24 h, subsequently infected with T. gondii for 24 h, and immunostained for ACTIN (red) and T. gondii GAP45 (green). The nucleus was stained with DAPI (blue). Arrow heads show vacuoles containing 2 or more parasites. Scale bars correspond to 5 μm. (D) WT, IDO1-KO or ATG16L1-KO HFFs, Huh7 or Hela cells were untreated or treated with IFN-γ for 24 h, and then infected with T. gondii. The number of parasites per vacuole after 24 h post infection was measured by IFA. Western blot and immunofluorescence images representative of three independent experiments (A,C). Indicated values are means of ± s.d. (three biological replicates per group from three independent experiments) (B,D). ***p < 0.001, **p < 0.01; N.S., not significant; (Student's t-test). N.D., not detected.