Toxoplasma gondii tachyzoites are intracellular and replicate in zebrafish. (A) Representative images from AiryScan confocal imaging of replicating tachyzoites in fixed larvae infected in the HBV with type I Toxoplasma-GFP at 6 and 24 hpi, showing 1, 2 or >4 tachyzoites/vacuole. Scale bar: 2 µm. (B) Pixel volume quantification of individual GFP-positive punctae at 6 and 24 hpi. Significant differences (χ22=58.5, ***P≤0.001) were observed between the percentage of total vacuoles counted in the HBV that are 1 tachyzoite/vacuole (<50 pix3), 2 tachyzoites/vacuole (50<100 pix3) or >4 tachyzoites/vacuole (>100 pix3) at 6 and 24 hpi. Pooled data from three independent experiments with at least three larvae per time point. Mean±s.e.m. shown. p.i., post-infection. (C) Representative AiryScan confocal images of replicating tachyzoites in larvae infected in the HBV with type I Toxoplasma-GFP (green; top-left images), fixed at 0, 6 and 24 hpi and labeled with α-GRA2 (red; top-right images) and merge (bottom large images). Shown are 1, 2 or >4 tachyzoites/vacuole. Scale bars: 2 µm.
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