File Name : sa1_ldh optimization licochalcone a.tif Caption : figure a1. optimization of the cell cytotoxicity assay in apical-in enteroids with licochalcone a as an example. in panels (a) and (b) cytotoxicity after 4 h and 24 h incubation and half-maximal cytotoxicity concentrations (ic₅₀) after 24 h incubation are shown without each enteroid dome treated as its own positive control, and in panels (c) and (d) with each enteroid dome treated as its own positive control. data are expressed as the mean ± sem of three biological replicates, measured in duplicate. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. File Name : sb2_influence passage number on ldh glabridin.tif Caption : figure b2. influence of passage number of mouse apical-in enteroids on glabridin cytotoxicity. panels (a) and (b) show glabridin cytotoxicity after 4 h and 24 h incubation in high (> 30) and low (< 13) passages, respectively, and panels (c) and (d) show half-maximal cytotoxicity concentrations (ic₅₀) of glabridin after 24 h incubation in high (> 30) and low passages (< 13), respectively. data are expressed as the mean ± sem of three biological replicates, measured in duplicate. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. File Name : sc3_gene expression markers passages.tif Caption : figure c3. gene expression of epithelial cell markers in apical-in and apical-out enteroids. panel (a) shows qrt-pcr gene expression analysis of epithelial cell markers for stem cells (lgr5), transit amplifying cells (ki67), goblet cells (muc2), enteroendocrine cells (chga), paneth cells (lyz), and enterocytes (villin) between low (p < 15, black dots) and high passages (p > 15, white dots) in apical-in enteroids grown in mouse (mic) and human (hic) growth medium, respectively, and apical-out enteroids (grown in hic). for the figures where the medium legend is not shown (e.g. mic and hic), these are similarly to those in the upper left figure. panel (b) shows the ratio between gene expression of epithelial markers (lgr5, ki67, muc2, chga, lyz) compared to villin gene expression in (1) jejunal scrapings from c57bl/6 mice¹ and apical-in enteroids grown in (2) mic and (3) hic, respectively. data are expressed as the mean ± sem of four biological replicates, measured in duplicate (pooled gene expression analysis of apical-in and apical-out organoids is shown in figure 5a). in vivo data from (b) were measured in three biological replicates. for the statistical analyses in (a) the pooled gene expression data was used (n = 8). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. File Name : sd4_toxicity and viability apical-out 24h.tif Caption : figure d4. cytotoxicity and cell viability of mouse apical-out enteroids. (a) enteroid cytotoxicity after 24 h glabridin incubation in µg ml^-1, (b) apical-out enteroid cell cytotoxicity after 24 h without prenylated phenolics incubation, and (c) cell viability at different time points without prenylated phenolics incubation. data in (a) are expressed as the mean ± sem of three biological replicates, measured in duplicate and in (b) and (c) are expressed as the mean ± sem of one independent biological replicate, measured in duplicate. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. File Name : sf5_biotransformation in apical-in organoids.tif Caption : figure f5. biotransformation of glabridin (glab, a), licochalcone a (licoa, b), and glycycoumarin (glycy, c) in apical-in mouse jejunal enteroids. sat-licoa = tentatively identified as saturated licochalcone a, glycy hydrated = tentatively identified as hydroxylation at prenyl group 3. identifications are shown in table e1. data are expressed as the mean ± sem of two biological replicates, measured in duplicate.