Journal: Nature
Article Title: Resolving the fibrotic niche of human liver cirrhosis at single cell level
doi: 10.1038/s41586-019-1631-3
Figure Lengend Snippet: a , Dotplot: ligand-receptor interactions between SAMΦ (n=10 human livers) and SAMes (n=7 human livers). X-axis, ligand (red) and cognate receptor (blue); y-axis, cell populations expressing ligand (red) and receptor (blue); circle size, P value (permutation test); colour (red, high; yellow, low), means of average ligand and receptor expression levels in interacting subpopulations. b , Representative immunofluorescence images, fibrotic niche. Left, TREM2 (red), PDGFB (white), PDGFRα (green), DAPI (blue), arrows TREM2 + PDGFB + cells. Right, TNFRSF12A (red), TNFSF12 (white), PDGFRα (green), DAPI (blue), arrows TNFRSF12A + PDGFRα + cells, scale bars 50μm. c to d , HSC proliferation assay: y-axis, area under curve (AUC) of % change in HSC number over time (hours), Mean±SEM, one-way ANOVA and Tukey. c , Control, TNFSF12, anti-TNFRSF12A, isotype control, all n=3. d , Vehicle, PDGF-BB, Crenolanib, all n=3. e , Dotplot: ligand-receptor interactions between SAEndo (n=7 human livers) and SAMes (n=7 human livers). X-axis, ligand (red) and cognate receptor (blue); y-axis, cell populations expressing ligand (red) and receptor (blue); circle size, P value (permutation test); colour (red, high; yellow, low), means of average ligand and receptor expression levels in interacting subpopulations. f , Representative immunofluorescence image, fibrotic niche. NOTCH3 (red), DLL4 (white), PDGFRα (green), DAPI (blue), arrows NOTCH3 + PDGFRα + cells, scale bar 50μm. g , Endothelial cell JAG1 flow cytometry: healthy (n=3) or cirrhotic (n=9) liver, representative histogram (left), mean fluorescence intensity (MFI, right), Mean±SEM, Mann-Whitney two-tailed. h , Cirrhotic endothelial cell and HSC co-culture, Notch inhibitor Dibenzazepine (DBZ). Representative immunofluorescence images (left), Collagen 1 (magenta), PECAM1 (green), DAPI (blue). Digital pixel analysis (right); collagen 1 area, n=3, Mean±SEM, RM one-way ANOVA and Tukey. n , HSC gene knockdown: control (n=7) or NOTCH3 (n=7) siRNA, qPCR of stated gene, expression relative to mean expression of control siRNA, Mean±SEM, Mann-Whitney two-tailed.
Article Snippet: After 2h, all growth factor supplements were removed and cells were cultured for a further 72 hours in endothelial basal media containing 10% heat-inactivated human serum ± Notch-signalling inhibitor Dibenzazepine (10μM, Bio-Techne, 4489/10) or vehicle (DMSO) control.
Techniques: Expressing, Immunofluorescence, Proliferation Assay, Flow Cytometry, Fluorescence, MANN-WHITNEY, Two Tailed Test, Co-Culture Assay
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