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Fig. 6.
Common human GBA1 mutants confer resistance to TB. (A) Bacterial burden measured by fluorescence in gba1sa1621/+ incross larvae 5 dpi after CV infection of 400 to 500 Mtb. Horizontal bars, means. *P < 0.05 (one-way ANOVA with Tukey’s posttest). (B) Schematic diagram showing the location of the two human GBA1 mutations studied. (C) Maximum intensity projection of representative pseudocolored confocal images of brain macrophages (yellow fluorescent) stained with LysoTracker Red in wild-type larvae, gba1 G0 crispants, and gba1 G0 crispants expressing human wild-type, L444P or N370S mutant GBA1 mRNA. (Scale bar, 20 μm.) (D) Mean LysoTracker Red volume per macrophage in 3 dpf animals from (C). Each point represents mean LysoTracker Red volume fraction per macrophage in each fish. Horizontal bars, means; ns, not significant; **P < 0.01 (one-way ANOVA with Tukey’s posttest). Representative of two to three independent experiments. (E) Bacterial burden measured by fluorescence in 5 dpi wild-type, gba1 G0 crispants and gba1 G0 crispants expressing human wild-type, L444P or N370S mutant GBA1 mRNA after CV infection with 600 to 700 Mtb. Horizontal bars, means; ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 (one-way ANOVA with Tukey’s posttest). Representative of two independent experiments. (F) Percentage of infected wild-type animals, gba1 G0 crispants and gba1 G0 crispants expressing human wild-type GBA1 mRNA fish at 5 dpi after HBV infection with a single Mtb bacterium. ns, not significant; *P < 0.05 (Fisher’s exact test). Representative of two independent experiments. (G) Percentage of infected wild-type animals, gba1 G0 crispants and gba1 G0 crispants expressing human L444P mutant GBA1 mRNA at 5 dpi after HBV infection with a single Mtb bacterium. *P < 0.05 and **P < 0.01 (Fisher’s exact test). (H) Percentage of infected wild-type animals, gba1 G0 crispants and gba1 G0 crispants expressing human N370S mutant GBA1 mRNA at 5 dpi after HBV infection with a single Mtb bacterium. **P < 0.01 (Fisher’s exact test). Representative of two independent experiments. (I) Total LysoTracker Red volume in 3 dpf wild-type, gba1 G0 crispants and gba1 G0 crispants expressing macrophage-specific human wild-type, L444P or N370S mutant GBA1 gene. Each point represents total LysoTracker Red volume in each fish. Horizontal bars, means. ns, not significant and *P < 0.05 (one-way ANOVA with Tukey’s posttest). (J) Bacterial burden measured by fluorescence in 5 dpi wild-type, gba1 G0 crispants and gba1 G0 crispants expressing macrophage-specific human wild-type, L444P or N370S mutant GBA1 gene after CV infection with 600 to 700 Mtb. Horizontal bars, means; ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001 (one-way ANOVA with Tukey’s posttest).