Figure 1. | Toxoplasma gondii F-actin forms an extensive filamentous network required for material exchange and parasite maturation

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Toxoplasma gondii F-actin forms an extensive filamentous network required for material exchange and parasite maturation

Figure 1.

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University of Glasgow, United Kingdom; Cancer Research United Kingdom Beatson Institute, United Kingdom; University of Vermont, United States
Figure 1.
Download figureOpen in new tabFigure 1. Analysis of intracellular development of act1 cKO.

(A) Conditional null mutants for act1 were induced as previously described in order to excise act1 (Egarter et al., 2014). 96 hr post induction, parasites were fixed and stained with anti-Gra2 to visualise the intravacuolar network. Scale bars: 10 µm. Replication assay of loxPact1 and act1 cKO. Number of nuclei per parasitophorous vacuole were counted 24 hr after inoculation on HFF cells. Mean values of three experiments in triplicate are shown. Asynchronous division is indicated by PVs with unusual amounts of parasites (≠2n). Two-way ANOVA followed by Sidak’s test were used to compare means between groups. ****p<0.0001, ***p<0.001, non-significance (ns) p>0.05. For source data see supplemental information (Figure 1A—source data 1). (B) Top image: The developing IMC of parasites is shown using IFA. Note that in case of act1 cKO individual parasites are in different stages of replication (i.e. white arrow indicates a parasite at the end of replication). Scale bars: 10 µm. Bottom image: Scanning EM analysis in intracellular parasites. Scale bars: 2 µm. Note the disordered appearance of parasites within the PV in absence of ACT1 and flattened bottom of the act1 KO parasites (Red Arrowhead). (C) Analysis of the IMC of the act1 cKO using IFA and scanning EM analysis in extracellular parasites. act1 cKO parasites have a flattened bottom, torpedo shape (Red arrowhead). The posterior pole of the parasite appears to be misformed, indicating a role of ACT1 during the final stages of replication (see also Figure 9). Scale bars: fluorescence images: 5 µm, SEM: 2 µm.

DOI: http://dx.doi.org/10.7554/eLife.24119.003