Extended Data Fig. 3: Responses to HEV infection in macrophages, organoids, and MaugOs.

a and b, Quantification of inflammatory gene expression in THP-1 macrophages (n = 4) or organoids (n = 4) upon HEV infection for indicated time points. Commentary to Fig. 3e. The exact p value * = 0.028. c, Schematic illustration of experimental design. Cholangiocyte organoids or hepatic differentiated organoids were first integrated with THP-1 macrophages for 24 hours to form MaugOs or Hep MaugOs, and then inoculated with HEV particles. Creaded in BioRender. van der Laan, L. (2025) https://BioRender.com/ulh914d. d, HEV infection kinetics in organoids, macrophages, or MaugOs (n = 6). e, Quantification of IL-1β and TNF-α cytokines in supernatant of organoids (ICO), macrophages (M), or MaugOs (n = 4). f, Brightfield and immunofluoresent imaging of hepatic differentiated organoids. Albumin (red): hepatocyte marker; DAPI (blue): nuclear staining. g, qRT-PCR quantification of the expression of stem cell marker (LGR5) and hepatic markers (albumin/ALB and glucose-6-phosphatase catalytic subunit/G6PC) in undifferentiated (ICO) and hepatic differentiated ICOs (HepO) (n = 4). The exact p value * = 0.028. h, Relative levels of HEV RNA (12 h normalized as 1) in hepatic differentiated organoids upon infection (n = 4). i, Quantification of HEV RNA level in MaugOs and Hep MaugOs 24 hours after HEV infection for 24 hours (n = 4). j, Production of IL-1β and TNF-α cytokines in supernatant of hepatic differentiated organoids (HepO), and Hep MaugOs upon HEV infection for 24 hours (n = 4). k and l, Relative levels of HEV RNA (12 h normalized as 1) and the expression of inflammatory genes quantified by qRT-PCR in HEV infected hiPSC-derived macrophages (n = 4). The exact p value * = 0.028. m, Quantification of HEV viral RNA in hepatic macrophages derived from perfusates of two donors inoculated with HEV for indicated time points (n = 4). Hepatic macrophages inoculated with HEV for 12 hours set as control (normalized as 1; each donor normalized to the corresponding control). n, Quantification of inflammatory gene expression in hepatic macrophages inoculated with HEV for 12 hours (three donors) and 24 hours (four donors). Uninfected groups served as control (normalized as 1). o, Quantification of inflammatory gene expression in HEV infected organoids integrated with hepatic macrophages (from two donors) for 12 or 24 hours. Uninfected MaugOs served as control (normalized as 1). (a, b, e, g, h, i, j k and l) Data are mean ± SD. *p < 0.05; **p < 0.01; Two-tailed Mann-Whitney U test. Exact p values are labelled, ‘n’ refers biological replicates.