41 2 Organoids as a model to study intestinal and liver dysfunction in severe malnutrition Figure 3. Amino acid deprivation induces barrier dysfunction in intestinal organoids. (A) Representative images for 10 kDa FITC-dextran for control (left) and starved organoid (right) (10x magnification). Scale bar, 100 μm (B) Average luminal FITC fluorescence for control, starved and rapamycin-administered organoids. Values are normalized to control organoids. Data represents mean of 3-4 biological replicates (from independent experiments) ± SEM (*P<0.05, two-way ANOVA with Tukey’s post hoc test). (C) Percentage of intestinal organoids with 4 or 10 kDa inside their lumen measured in organoids grown in complete culture medium (control), amino-acid-free medium (starved) and amino-acid-free medium supplemented with 2nM rapamycin (rapamycin). Data represents mean of 3 biological replicates (from independent experiments) ± SEM (*P<0.05, two-way ANOVA with Tukey’s post hoc test). (D) Immunoblot quantification and representative image of tight-junction protein claudin-3 relative to total protein (see methods) for the different conditions. Normalized to total protein. Data represents mean of 3 biological replicates (from independent experiments) ± SEM. (*P<0.05, ** P< 0.01, Two-way ANOVA with Tukey’s post-hoc test). (E) Relative intensity of total protein, summed over the entire lane (used for normalization to total protein, as explained in methods). Individual data points are shown for each biological replicate.
RkJQdWJsaXNoZXIy MTk4NDMw