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. 2016 Oct 10;17(12):1753–1765. doi: 10.15252/embr.201642358

Figure 4. Rab22a is recruited to the parasitophorous vacuole of Toxoplasma gondii and is necessary for cross‐presentation of a parasite‐derived soluble antigen.

Figure 4

  1. Scramble and Rab22a KD JAWS‐II DCs were infected with OVA‐YFP‐expressing T. gondii (TgRH YFP SAG1‐OVA) for 8 h and confocal images detecting the parasite (green), endogenous Rab22a (red), and GRA6 (magenta) were taken. Top panels: Scramble cells; bottom panels: Rab22a KD cells. White boxes are shown at higher magnification in the insets. The nuclear marker DAPI (blue) and DIC images are shown in the left panels. Overlays are shown in the right panels. Scale bars: 5 μm. Images are representative of at least 30 analyzed from three independent experiments.
  2. The cross‐presentation of OVA secreted by T. gondii (TgRH YFP SAG1‐OVA) after 8 h of infection at the indicated MOI was evaluated by B3Z T‐cell activation. Data represent mean ± SEM of triplicate values and are representative of three independent experiments. ***P < 0.001. A two‐way ANOVA and the Bonferroni post‐test were performed.
  3. The efficiency of infection (8 h) of TgRH YFP SAG1‐OVA in Scramble and Rab22a KD JAWS‐II DCs was measured by FACS analysis at the indicated MOI. Data represent mean ± SEM of duplicate values and are representative of three independent experiments.
  4. Immunoblotting showing the total amount of OVA in Scramble and Rab22a KD JAWS‐II DCs after 90 min and 7 h of infection with TgRH YFP SAG1‐OVA parasite at MOI 2 and 6. Fifty micrograms of total cell lysates was loaded onto each lane. Data are representative of two independent experiments.

Source data are available online for this figure.