Journal: Gastro Hep Advances
Article Title: Proliferative Cell Targeting and Epithelial Cell Turnover Fuels Hepatitis E Virus Replication in Human Intestinal Enteroids
doi: 10.1016/j.gastha.2025.100769
Figure Lengend Snippet: Targeted cell tropism of HEV in infected HIEs. (A) Percentage of proliferative cells in mock-, HEV-3 or HEV-3 G1634R -electroporated fetal ileum HIEs at day 1, 7, 11 and 15 pe. Amount of proliferative cells was determined by HCI and defined by the objects positive for both DAPI and the proliferation marker Ki67. (B) Representative image of immunofluorescence staining of HEV-3 G1634R electroporated fetal ileum HIEs at day 11 pe. Cells were stained with HEV ORF2 (green) and Ki67 (red). (C) Percentage of HEV ORF2 capsid positive cells within the proliferative cell population in mock-, HEV-3 or HEV-3 G1634R -electroporated fetal ileum HIEs at day 1, 7, 11 and 15 pe. These cells were defined by the objects positive for all three markers (Ki67 and HEV ORF2). (D) Percentage of cells that were positive for proliferative marker (Ki67) within the HEV ORF2 positive population in mock-, HEV-3 or HEV-3 G1634R electroporated fetal ileum HIEs at day 1, 7, 11 and 15 pe. These cells were defined by the objects positive for all three markers (DAPI, Ki67 and HEV ORF2). (E, F) Representative images of immunofluorescence staining of HEV-3 G1634R electroporated fetal ileum HIEs at day 11 pe. Cells were stained with HEV ORF2 (green) and antibody specific for (E) enteroendocrine cells (CHGA, chromogranin A, red) or (F) goblet cells (WGA, wheat germ agglutinin). DAPI (blue) was used as counterstaining. Overlap of voxels between HEV ORF2 and (E) CHGA signal ( N = 3, 8 fields/experiment) or (F) WGA signal) ( N = 3, 5 fields/experiment). pe, postelectroporation.
Article Snippet: The following antibodies or dyes were used: anti-ORF2 (HEV-specific rabbit hyperimmune serum), anti-Ki67 (EMD Millipore, MAB4190), antisucrose isomaltase (Santa Cruz Biotechonology, sc-393424), anti-CHGA (Santa Cruz Biotechonology, sc-393941), phalloidin (Invitrogen, A12380) or wheat germ agglutinin (WGA, Vector laboratories, RL-1022).
Techniques: Infection, Marker, Immunofluorescence, Staining
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