Fig. 6: Hyperplasia and hypertrophy of midlobular hepatocytes repair the liver after coronavirus injury. | Nature Communications

Fig. 6: Hyperplasia and hypertrophy of midlobular hepatocytes repair the liver after coronavirus injury.

From: Heterogeneity of hepatocyte dynamics restores liver architecture after chemical, physical or viral damage

Fig. 6

A Schematic showing the experimental design for coronavirus liver injury with MHV-A59. Created with BioRender.com. B Body weight curves in 10-week-old male mice during the first week of viral infection with MHV-A59 or PBS vehicle treatment; n = 3–4 mice per group. Data are presented as mean values +/− SEM. C Liver sections from mice after 1 and 4 weeks of MHV-A59 viral infection. Asteriks indicate immune cell infiltrates. CV central vein, PV portal vein. Scale bars, 100 μm. D Quantification of DAPI staining or total number of cells per field of view (10×) in liver sections from mice before and after 1 and 4 weeks of MHV-A59 infection. Data are presented as mean values +/− SEM. 5–8 photos analyzed per mouse; n = 3 mice per group. One-way ANOVA, Tukey’s multiple comparisons post-test. E Quantification of CAG-EGFP staining per field of view (10×) in liver sections from mice 1 and 4 weeks post-MHV-A59 infection. Data are presented as mean values +/− SEM. 5–8 photos analyzed per mouse; n = 3 mice per group. One-way ANOVA, Tukey’s multiple comparisons post-test. F Hepatocyte injury 1 week post-MHV-A59 infection in livers from mice. Left: Relative frequency in % of each clone size per area of the liver lobule and per Rainbow fluorophore from mice at 1 week post-MHV-A59 infection. Data are presented as mean values of the average PP, Mid and PC hepatocyte population +/− SEM. 6 liver lobule areas analyzed per mouse; 2 mice per time-point (4–5 photos per mouse). Middle: quantification of number of cells per clone (y-axis) and number of clones (x-axis) per area of the liver lobule and per Rainbow fluorophore from mice at 1 week post-MHV-A59 infection. Data are presented as mean values. 6 liver lobule areas analyzed from 2 mice per time point (4–5 photos per mouse). One-way ANOVA, Tukey’s multiple comparisons post-test. Right: Representative images of a liver lobule from mice at 1 week post-MHV-A59 infection; n = 2 mice. G Clonal expansion of hepatocytes 4 weeks post-MHV-A59 infection in livers from mice. Left: Relative frequency in % of each clone size per area of the liver lobule and per Rainbow fluorophore from mice at 4 weeks post-MHV-A59 infection. Data are presented as mean values of the average PP, Mid and PC hepatocyte population +/− SEM. 6 liver lobule areas analyzed per mouse; 2 mice per time-point (4–5 photos per mouse). Middle: quantification of number of cells per clone (y-axis) and number of clones (x-axis) per area of the liver lobule and per Rainbow fluorophore from mice at 4 weeks post-MHV-A59 infection. Data are presented as mean values. 6 liver lobule areas analyzed per mouse; 2 mice per time point (4–5 photos per mouse). One-way ANOVA, Tukey’s multiple comparisons post-test. Right: Representative images of a liver lobule from mice at 4 weeks post-MHV-A59 infection; n = 2 mice. Scale bars, 100 μm. H Quantification of proliferating Ki67 hepatocytes per field of view (10×) in each zone of the liver lobule from mice at week 4 post-MHV-A59 infection; Data are presented as mean values +/− SEM. 9–15 liver lobule areas analyzed from 2 mice per time point. One-way ANOVA, Tukey’s multiple comparisons post-test. I Hepatocyte hypertrophy after 4 weeks of MHV-A59 infection. Left: Hepatocyte size after tissue repair in liver sections from mice 4 weeks after MHV-A59 infection. Data are presented as mean values +/− SEM. Hepatocyte size (n = 53–91), from 3–4 liver lobule areas were analyzed from 2 mice. Unpaired Student’s t-test. Right: Hepatocyte size (n = 29) within the liver lobule from mice at 4 weeks after MHV-A59 infection. Data are presented as mean values +/− SEM. 3–4 liver lobule areas analyzed from 2 mice. One-way ANOVA, Tukey’s multiple comparisons post-test. (J) Representative CK19 IHC in liver sections from mice at 1 and 4 weeks after MHV-A59 infection. Arrows indicate cholangiocytes forming bile ducts around the portal triads; n = 2 mice. K Hepatocyte ploidy in the liver lobule of mice at 4 weeks after MHV-A59 infection. % of total polyploid and diploid hepatocytes within the liver lobule (left graph) and per area of the liver lobule (middle left graph). % of diploid hepatocytes (hepatocytes that express only 1 fluorophore) being mononucleated or multinucleated (middle right graph). % of polyploid hepatocytes (hepatocytes that express more than 1 fluorophore) being mononucleated or multinucleated (right graph). Data are presented as mean values +/− SEM. 5 photos analyzed per mouse; 2 mice per group. Source data are provided as a Source Data file.

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