Fig. 5: Proliferation of commensal L. jeotgali LUAb3 and pathogenic Ca. L. limosiae LUAb1 bacteria in C. elegans N2 gut lumen. | npj Biofilms and Microbiomes

Fig. 5: Proliferation of commensal L. jeotgali LUAb3 and pathogenic Ca. L. limosiae LUAb1 bacteria in C. elegans N2 gut lumen.

From: Dynamics of gut colonization by commensal and pathogenic bacteria that attach to the intestinal epithelium

Fig. 5

a-c Representative RNA FISH-stained images of C. elegans N2 briefly exposed to LUAb3 and fixed 0 h post exposure (hpe), 24 hpe, and 48 hpe respectively. Red fluorescence correlates with LUAb3 specific RNA FISH probes with a Cal-610 fluorophore. Scale bars are 100 µm. d Quantification of fluorescence intensity of LUAb3 specific fluorescent probes for each fixed time point. n = 30 in 3 independent experiments. Statistical analysis performed by Tukey’s multiple comparisons test in a one-way ANOVA where **p < 0.05 and ****p < 0.001. e Percentage of C. elegans N2 colonized with LUAb3 in the intestine at 24 hpe and 48 hpe (N = 3, n = 30). f Quantification of the colony forming units (CFUs) per L. jeotgali LUAb3 or L. amnigena JUb66a colonized animal up to 48 hpe. Data is normalized to animals exposed to only E. coli OP50-1 and consist of two independent experiments performed in biological triplicate. Statistical analysis performed by Tukey’s multiple comparisons test in a One-Way ANOVA where ****p < 0.0001 and ns = not significant. g Categories of directional LUAb1 (red fluorescence) colonization in C. elegans, where 0 represents no colonization, 1 represents LUAb1 present only in the anterior end, 2 is LUAb1 detected between the anterior and midline of the animal (vulva), 3 is LUAb1 found from the anterior end past the midline, and 4 is LUAb1 colonizing the entire anterior-posterior length of the intestine. Scale bars are 100 µm. h–j Representative RNA FISH-stained images of C. elegans N2 exposed to LUAb1 briefly and fixed at 24 hpe, 48 hpe, and 72 hpe. Scale bars are 100 µm. k Percent of LUAb1 colonized animals in each biogeographical category for each time point. l Quantification of the percentage of animals colonized past the midline (categories 3 and 4) in each time point. n is at least 90 animals in two independent experiments. Statistical analysis performed by Tukey’s multiple comparisons test in a One-Way ANOVA where *p < 0.05, **p < 0.005, and ns = not significant.

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