Extended Data Fig. 5: FXR modulation in biliary, airway and intestinal organoids.
From: FXR inhibition may protect from SARS-CoV-2 infection by reducing ACE2
(a) QPCR analysis in primary airway, biliary and intestinal organoids demonstrating that CDCA activates FXR, whereas UDCA and ZGG inhibit it, as evidenced by corresponding changes in the expression of the FXR downstream target SHP. Housekeeping gene, HMBS; n = 4 independent experiments; one-way ANOVA adjusted for multiple comparisons; centre line, median; box, interquartile range (IQR); whiskers, range; bars, standard deviation. (b) Immunofluorescence showing ACE2 expression levels in primary organoids in absence of bile acids. The panel is complementary to Fig. 1d showing the modulation of ACE2 following FXR activation (CDCA) and inhibition (UDCA/ZGG). N = 4 independent experiments. Yellow scale bars 50 μm; grey scale bar 25 μm. (c) Flow cytometry histograms showing changes in ACE2 levels upon modulation of FXR activity in primary airway, biliary and intestinal organoids. n = 3 independent experiments. (CDCA, UDCA and ZGG concentration, 10 μM). (d) Dose–response curves showing the effect of 0.01 μM – 1 mM of CDCA, UDCA and ZGG on the expression of ACE2 in primary airway, biliary and intestinal organoids (n = 3 independent experiments). Response defined as percentage of the maximal ACE2 expression level for each condition via QPCR. Bars, SEM. (e) Percentage of non-viable cells following treatment of airway, biliary and intestinal organoids with CDCA, UDCA and ZGG at a range of 0.1 μM – 100 μM showing that these compounds cause minimal cell death within the tested range. N = 3 independent experiments. Bars, range. (f) Resazurin assay (PrestoBlue) showing that treatment with 10 μM of CDCA, UDCA or ZGG does not affect cellular viability. N = 4 independent experiments; one-way ANOVA adjusted for multiple comparisons; ns, non-significant; centre line, median; box, interquartile range (IQR); whiskers, range; bars, standard deviation.
