Data table of the Pathogen Box screen for inhibitors of mitochondrial O2 consumption in T. gondii parasites.

(Tab 1A-B) Plate configurations of the two Seahorse XFe96 plates used in the screen. Included are the MMV compounds that were injected into the indicated wells from Ports A-C during the first (Tab 1A) and second (Tab 1B) assays. Hit compounds are highlighted in yellow, compounds excluded from subsequent analyses because they were injected after hit compounds are highlighted in blue, compounds rescreened on the second plate are highlighted in green, background wells are highlighted in orange, and wells from a separate experiment that was included on Plate 2 are indicated in gray. Atovaquone and antimycin A was injected from Port D into all wells. (Tab 2A-B) Calculated OCR and ECAR values obtained for each well at each measurement time in Plate 1 (Tab 2A) and Plate 2 (Tab 2B). The time from the commencement of the assay at which each measurement was taken is indicated. Injection of compounds from Port A occurred between the third and fourth measurements, injection from Port B occurred between the seventh and eighth measurements, injection from Port C occurred between the 11th and 12th measurements, and injection from Port D occurred between the 15th and 16th measurements. Also indicated are the average OCR and ECAR values across all wells at each measurement point, with the average OCR value in the final measurement following atovaquone/antimycin A injection used to determine the non-mitochondrial OCR for the assay (highlighted in yellow). (Tab 3) Summary of the assay data, expressed in a table modified from the Pathogen Box plate mapping spreadsheet provided by MMV. Included are: the Pathogen Box plate and position of the test compounds; the MMV compound identification number and common name (where applicable); the Seahorse XFe96 plate well position and injection port from which the compound was injected during the assay (with compounds injected into Plate 1 indicated in green and compounds injected into Plate 2 in yellow); the mitochondrial OCR (mOCR) values immediately before and after compound injection (calculated by subtracting the non-mitochondrial OCR from the OCR values listed in Tab 2A-B; note that the post-drug values were obtained on the fourth measurement, approximately 18 min, after compound injection for all compounds except auranofin, for which we noticed a more gradual OCR decrease, and which was therefore calculated after ~40 min); the percent inhibition of mitochondrial OCR following compound injection (calculated from the difference between pre-compound injection and post-compound injection mOCR values); an indication of whether compounds met the >30% inhibition cut-off for hit compounds; and the chemical structures of all MMV compounds expressed in simplified molecular input line entry system (SMILES) notation. (Tab 4A-B) The O2 and pH values obtained during each measurement in the Seahorse XFe96 assays in plate 1 (Tab 4A) and plate 2 (Tab 4B). These values form the basis from which the OCR and ECAR values depicted in Tab 2A-B were determined. (Tab 5A-B) Calibration data for the OCR (left) and ECAR (right) assays conducted in plate 1 (Tab 5A) or plate 2 (Tab 5B). Note that the pH calibrations failed for some wells in plate 1 and we therefore did not include ECAR data in our analysis. (XLSX)