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. 2014 Apr 29;12(4):e1001845. doi: 10.1371/journal.pbio.1001845

Figure 4. MAF1 (TGGT1_053770) mediates Toxoplasma HMA.

Figure 4

(A) Type II or type II parasites expressing an N-terminally HA-tagged TGGT1_053770 transgene were used to infect HFFs. Cells were fixed 12 hpi, and following permeabilization, TGGT1_053770 was visualized by immunofluorescence microscopy using polyclonal mouse sera raised against recombinant TGGT1_053770; host mitochondria were visualized using rabbit anti-TOM20 antibodies. (B) Transmission electron micrographs depicting the PVM surrounding type II and type II:TGGT1_053770 parasites grown in mBMDMs. Cells were fixed and processed for electron microscopy 6 hpi. Host mitochondria are indicated by M. (C) Percentage of the PVM associated with mitochondria in HFFs 4 hpi with type I and type I:Δmaf1, and BMMs 17 hpi with type II and type II:MAF1 vacuoles as determined by ImageJ analysis of electron micrographs (n = 20 for each). Values shown are mean ± SEM. ****p<0.0001 using an unpaired t test. (D) Type I and type I:Δmaf1 parasites (expressing RFP) were used to infect HFFs, which were then fixed 4 hpi. Following permeabilization, host mitochondria were visualized using rabbit anti-TOM20 antibodies. (E) Transmission electron micrographs depicting the PVM surrounding type I and type I:Δmaf1 parasites grown in HFFs. Cells were fixed and processed for electron microscopy 4 hpi. Host mitochondria indicated by M. Scale bars are 5 µm (A and C) and 0.5 µm (B and D).