Figure 4.

Active GULO carrying eHLOs conjugate bilirubin more efficiently

(A) Bright-field and fluorescence images of mCherry expression in Dox-treated mGULO HLOs compared with control HLOs. Scale bar, 200 μm.

(B) Bright-field image of mGULO HLOs treated with bilirubin (10 mg/L) and Dox (0–1,000 ng/mL) (top). UnaG assay for mGULO organoids treated with Dox compared with control (bottom). Scale bars, 200 μm.

(C) Bilirubin assay measuring unconjugated and conjugated bilirubin in mGULO HLOs treated with bilirubin (10 mg/L) and Dox (0–1,000 ng/mL). n = 9 independent experiments.

(D) UnaG assay shows conjugation of bilirubin indicated by loss of fluorescence even in the presence of dark yellow bilirubin. Scale bar, 200 μm.

(E) qRT-PCR of NRF2 and IL6 gene for mGULO organoids treated with Dox (100 ng/mL) compared with control after treatment with bilirubin. Data are mean ± SD, n = 9 independent experiments.

(F) Total percentage of viable organoids and organoids carrying conjugated bilirubin in Dox-treated organoids compared with control. n = 9 independent experiments.

ns, p > 0.05, ^∗p ≤ 0.05, ^∗∗p ≤ 0.01, ^∗∗∗p ≤ 0.001, and ^∗∗∗∗p ≤ 0.0001.