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. 2024 May 23;15:4385. doi: 10.1038/s41467-024-48685-4

Fig. 2. F97L mutation in eIF1.2 led to a differentiation defect in vitro and low cyst burden in vivo.

Fig. 2

a eIF1.2 F97L mutant. b–d Plaque assay. Mean ± s.d. for 6 biological replicates. NS, not significant (P > 0.05); Student’s two-tailed t-test. e–o Parasites exposed for 7 days to alkaline stress. e–j Flow cytometry analysis and quantification of tdTomato(+) and GFP(+) cell percentage (f, i) and intensity (g, j) in total ungated population. Numbers in (e) and (h) represent the percentage of the gated population relative to the total population. Data conducted on the same day were connected by a line. AU, arbitrary units. n = 5 biological replicates; NS, not significant (P > 0.05); Student’s two-tailed t-test. k Phase contrast microscopy images. Bar, 50 μm. l In vitro cysts count. Mean ± s.d. for n = 3 biological replicates; P-values derived from a Poisson model. m Immunofluorescence images. Bar, 20 µm. n, o Quantification of DBA and GFP signals within individual vacuoles. Mean ± s.d. for at least n = 300 vacuoles from 3 biological replicates. Data were analyzed with a linear regression model. p–r Western blot analysis. Stressed, 7 days of alkaline stress. Actin, loading control. Mean ± s.d for 4 biological replicates. Data were analyzed using a linear mixed model. NS, not significant (P > 0.05). s Bioluminescence imaging. Uninfected (n = 2 mice), WT (n = 8 mice), eIF1.2 F97L (n = 8 mice). Data represent mean ± s.e.m. and analyzed using a linear regression model. t Mice (s) body weight. Uninfected (n = 2 mice), WT (n = 8 mice), eIF1.2 F97L (n = 8 mice). Data represent mean ± s.d. and were analyzed using a linear regression model. NS, not significant (P > 0.05). u Mice survival curve. n = 12 mice for each group. NS, not significant (P > 0.05); Mantel-Cox two-sided test. v qPCR analysis of parasite burden in mice brain. WT (n = 10 mice), eIF1.2 F97L (n = 9 mice). Data represent mean ± s.d. and analyzed using Welch’s two-tailed t-test. Source data are provided as a Source Data file.