Journal: bioRxiv
Article Title: Human enterochromaffin cells and apical-out intestinal organoids as models for human sapovirus infection
doi: 10.64898/2026.03.06.710023
Figure Lengend Snippet: Phylogenetic tree of sapovirus samples (n = 27), color coded by genotype, and reference strains. The tree was constructed with Maximun likelihood algorithm using Kimura 2-parameter model (gamma distributed with invariant sites) and 1000 bootstrap replicates. Scale bar for branch lengths shows number of substitutions per site. Samples F20, F21, F30 and F31 were excluded in the analysis due to incomplete sequences. Reference strains: GII.1/Hu/IND/2021/NICED-BCH-12967 (GenBank: LC851686 ), GII.1/Hu/JPN/2013/Mie-493 (GenBank: LC816490 ), GII.1/Hu/BGD/2012/475050 (GenBank: MW088928 ), GII.1/Hu/UK/1998/Bristol (GenBank: AJ249939 ), GII.3/Hu/JPN/2023/20220111 (GenBank: LC848995 ), GII.3/Hu/JPN/2023/S22-415 (GenBank: LC849000 ), GII.3/Hu/JPN/2019/Mie-021 (GenBank: LC816544 ), GII.3/Hu/JPN/2017/Mie-835 (GenBank: LC816541 ), GI.1/Hu/US/2014/ Portland3639 (Genbank: MG012399 ), GI.1/Hu/IND/2022/NICED-BCH-13235 (GenBank: LC851695 ), GI.1/Hu/JPN/2011/EH-70 (GenBank: LC504316 ), GI.1/Hu/JPN/2023/V231401 (GenBank: LC848977 ), GI.1/Hu/JPN/1982/MT-2010 (GenBank: HM002617 ), GII.1/Hu/US/2015/Nashville9343 (GenBank: MG012444 ), GI.1/Hu/GER/pJG-Sap01/Dresden (GenBank: AY694184 ), GI.3/Hu/SWE/1997/318/Stockholm (GenBank: AF194182 ), GII.3/Hu/US/2015/Nashville9354 (GenBank: MG012419 ), GI.1/Hu/UK/1993/Manchester (GenBank: X86560 ). Abbreviations: Hu: Human, IND: India, JPN: Japan, US: United States, BGD: Bangladesh, GER: Germany, SWE: Sweden, UK: United Kingdom. B) Viral load (genome copies/gram of feces) and genotype of 31 sapovirus positive samples. Each colored square corresponds to one sample, and the median viral load is shown for each genotype. p-value, determined by Kruskal-Wallis test, is displayed. C) Detection of CD36 by gene expresison analysis and immunofluorescence staining in GOT1 and HuTu80. Gene expression of CD36 is displayed as inverted ΔCt value (1/ ΔCt), with ΔCt calculated by subtracting the Ct-value for CD36 to the Ct of the house-keeping gene GAPDH. The fold difference in expression between HuTu80 and GOT1 is shown above the bars. D - F) Replication of sapovirus samples in HuTu80 and GOT1. Left y-axis represents the fold change (FC) of virus at 72 hours post infection (hpi) compared to 2 hpi. The two biological duplicates per infection experiment are shown as black dots. A mean FC of at least 5-fold (dotted line) was considered successful replication and samples with a mean FC of less than 1 were set to 1.1 FC for visualization purposes in the graph. The sample viral load, expressed as genome copies per gram of feces, is shown as colored squares. GAPDH; Glyceraldehyd-3-fosfatdehydrogenas
Article Snippet: Also, since HuTu80 cells originate from EECs, a duodenal adenocarcinoma of L-cell type (ATCC HTB-40), we investigated whether another EEC cell line, GOT1, could serve as a potential model for sapovirus infection.
Techniques: Construct, Immunofluorescence, Staining, Gene Expression, Expressing, Virus, Infection