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. 2020 Oct 20;11(5):e02092-20. doi: 10.1128/mBio.02092-20

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Copyright © 2020 Bisio et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 7 — TgStx12 is essential for microneme and rhoptry function. (A) Indirect immunofluorescence assay (IFA) showing the endogenously N-terminally tagged Stx12 localized to the ELC. ProM2AP, parasite ELC marker. (B) Rapamycin-induced Stx12 downregulation in intracellular parasites within 24 hours. GAP45, parasite pellicle. (C) The Stx12-LoxP-U1 knockdown but not the parental DiCre strain displays a severe growth defect in the presence of rapamycin. (D) Parasites lacking Stx12 show a significantly impaired invasion capacity. Error bars represent the ± SD for three independent experiments. (E) The content of microneme and rhoptry localized proteins is reduced upon depletion of Stx12. Catalase, loading control. (F) The quantification of RON4 and MIC2 are shown. Error bars represent the ± SD for three independent experiments. (G) Electron microscopy reveals no gross morphological defects of micronemes and mild abnormalities of rhoptries in parasites depleted in Stx12. Micronemes and rhoptries are indicated with red and white arrowheads, respectively. Scale bars = 1 μm. Scale bars for all IFA = 7 μm.