Fig. 8: Pantothenate synthesis is essential for chronic infection of T. gondii in vivo. | Nature Communications

Fig. 8: Pantothenate synthesis is essential for chronic infection of T. gondii in vivo.

From: Pantothenate biosynthesis is critical for chronic infection by the neurotropic parasite Toxoplasma gondii

Fig. 8

a Scheme of the Pan biosynthesis pathway, highlighting the ketopantoate hydroxymethyl transferase - ketopantoate reductase (KPHMT-KPR) and the pantoate beta alanine ligase (PBAL). See Fig. 1 for abbreviations. b Survival curve of mice infected with RH parental (n = 10), kphmt-kpr-ko (n = 5) and pbal-ko (n = 5) parasites (from 2 experiments, logrank Mantel–Cox test). c Western blot of complementant strains pbal-ko::cTgPBAL-myc (n = 3) and pbal-ko::cEcPBAL-myc (n = 4). Antibodies anti myc, anti actin as loading control. d IFA of pbal-ko::cTgPBAL-myc (n = 5) and pbal-ko::cEcPBAL-myc (n = 3). Antibodies anti myc, anti GAP45 as marker of parasite pellicle. e Plaque assay of ME49 parental, pbal-ko and complement strains::cTgPBAL-myc,::cEcPBAL-myc (n = 3). f Quantification of plaque size. Black dots present mean values of independent experiments (n = 3). g Survival curves of mice infected with ME49 parental (n = 41, 24 survived), ME49 kphmt-kpr-ko (n = 12, 11 survived), ME49 pbal-ko (n = 17, 17 survived), ME49 pbal-ko::cTgPBAL-myc (n = 8, 7 survived), ME49 pbal-ko::cEcPBAL-myc (n = 8, 7 survived) strains, pooled from 5 independent experiments. h Ex vivo cyst count from mouse brains infected for 5 weeks with parental ME49, kphmt-kpr-ko, pbal-ko, pbal-ko::cTgPBAL-myc, and pbal-ko::cEcPBAL-myc parasites (n = 25, 9,15, 7 and 7, respectively, pooled from 5 independent experiments). Empty circles present counts before the 5 weeks endpoint. Median is represented, 2-sided Mann–Whitney test. i Representative plaque assay of brain-derived cysts from ME49 parental, kphmt-kpr-ko and pbal-ko infected mice (n = 3, 3, 4, respectively). j Cartoon schematic depicting the different modes of Pan acquisition in tachyzoites (Tz) and bradyzoites (Bz). n number of independent biological replicates. Source data are provided as a Source Data file. White scale bar 5 µm. Black scale bar 1 mm. Red asterisk marks degradation product of tagged protein. Black asterisk marks unspecific signal.

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