68 Chapter 2 Supplementary Figure 6. FITC fluorescence in the lumen of intestinal organoids. FITC fluorescence was measured in the lumen of intestinal organoids that were grown in complete culture medium (control), amino-acid-free medium (starved) and amino-acid-free medium supplemented with 2nM rapamycin (rapamycin). Graphs represent three different organoid lines. Data points are individual organoids random selected by a random generator and the error bars indicate the SEM. (*P <0.05, *** P<0.001, two-way ANOVA with Tukey’s post hoc test). Supplementary Table 1. Amino acid concentrations measured in the medium of mature hepatic organoids. Concentration (µM) Amino acid Control medium Amino-acid-starvation medium Alanine 1405 ± 110 86 + 19 Asparagine 32 ± 8 7.0 ± 0.2 Glutamic Acid 136 ± 169 166 ± 233 Glutamine 480 ± 164 10 ± 1 Glycine 144 ± 3 5 ± 0.1 Isoleucine 328 ± 11 8 ± 2 Leucine 327 ± 9 10 ± 2 Lysine 284 ± 26 16 ± 4 Phenylalanine 188 ± 10 10 ± 2 Proline 162 ± 22 41 ± 5 Serine 147 ± 9 2 ± 0 Tyrosine 151 ± 10 14 ± 1 Valine 410 ± 3 18 ± 4 Amino acid concentrations were measured in medium that had been in the well with organoids for 48 hours. Data is expressed as mean ± SD and represents 3 biological replicates (from independent experiments) for control medium and 2 biological replicates for amino-acid-starvation medium measured in hepatic organoid cultures.
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