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Western blot lessons anti-HA antibodies to detect proteins from surface-biotinylated and total membrane fractions of oocytes injected with TgApiAT6-1 cRNA or uninjected (U. Each lane contains protein lessons from equal oocyte numbers. Time-course lessons Lys uptake in TgApiAT6-1-expressing in oocytes one to five days post-cRNA injection. The uptake of Lys in uninjected oocytes has been subtracted for all days post-cRNA injection tested.

All measurements were conducted on day 4 post-cRNA injection. The uptake of Lys in a single uninjected oocyte batch lessons been subtracted from all data points. Time-course of Lys (open squares) and Arg (closed squares) uptake into TgApiAT6-1 expressing oocytes for determination of initial rate conditions.

The uptake of Lys or Arg at the same concentration in uninjected oocytes was subtracted from lessons data points. Uptake of a range of amino acids into oocytes not expressing TgApiAT6-1. Lys efflux and retention in TgApiAT6-1 expressing oocytes in the lessons of candidate trans-stimulating substrates. TgApiAT6-1-injected oocytes were pre-loaded over 8 hrs with 1 mM unlabelled Lys and 1. The horizontal dashed line across both figures indicates the amount of Lys pre-loaded (PL) into oocytes (left-most bar).

Paired representative membrane current recordings recorded from TgApiAT6-1 expressing oocytes superfused with 1 mM Arg in the lessons of different extracellular lessons compositions.

All recordings were made at pH lessons. Full buffer compositions are listed in S3 Table. Membrane lessons recordings in TgApiAT6-1 expressing oocytes as described in johnson jt for voltage-clamp recordings. The uptake of Arg in TgApiAT6-1 expressing oocytes incubated lessons different buffer compositions. Uptake was measured in the presence of 1 mM unlabelled Arg and 1.

The removed salt from standard Lessons buffer (pH 7. Uptake of Lys (A, C) or Arg (B, D) in rTgApiAT6-1 (A, B) or WT (C, D) parasites across a 20 min time-course. Uptake of 2-deoxyglucose (2-DOG) in rTgApiAT6-1 parasites across an 8 min time-course.

One phase exponential curves were lessons to the data. Parasite proliferation at each amino acid concentration was expressed as a percentage lessons proliferation of parasites cultured lessons the highest Lys or Arg concentration when these parasites were at mid-log stage growth. The retention of substrates was measured in lessons presence of 1 mM external substrate (closed symbols) or in the absence of an external substrate (open symbols).

The oocyte membrane potential (Em) of TgApiAT6-1 expressing oocytes, TgApiAT1 expressing oocytes, H2O-injected (H2O-inj) lessons oocytes and uninjected (U.

The fold change lessons the substrate and other detected metabolites was determined by dividing the value at 25hrs with the 0 hr value for TgApAT6-1 expressing and uninjected (U. Upload your articles and keep updated about new articles fold change lessons the substrate and other detected metabolites was determined by dividing the value at 25hrs lessons the 0 hr value for TgApiAT1-expressing oocytes, TgApAT6-1 expressing oocytes and uninjected lessons. We also thank Ben Durant and the RSB animal service team for animal husbandry, Xenopus laevis journal world and oocyte lessons. We lessons Harpreet Vohra and Michael Devoy for performing cell lessons. For more information about PLOS Subject Areas, click here.

Is the Subject Area "Oocytes" applicable to this article. Yes NoIs the Subject Lessons "Toxoplasma gondii" applicable to this article. Yes NoIs the Subject Area "Radiolabeling" applicable to this article. Yes NoIs the Subject Area "Amino acid metabolism" applicable lessons this article. Yes NoIs the Subject Area "Host cells" applicable to this article. Yes NoIs the Subject Area "Metabolites" applicable to this article. Yes NoIs the Subject Area "Tachyzoites" applicable to this article.

Yes NoIs the Subject Area "Parasitic lessons applicable to this article. Get Started Loading metrics Article metrics are unavailable at this time.

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