Extended Data Fig. 6: Analysis of the myHSC and cyHSC subpopulations in mouse and human fibrotic livers.
From: Opposing roles of hepatic stellate cell subpopulations in hepatocarcinogenesis
a, UMAPs of myHSC and cyHSC signatures by scRNA-seq, each visualized specific signatures as well as the correlation of cyHSC and myHSC signature score in HSC isolated from mice fed with HF-CDAA NASH diet for 12 weeks (n = 1 mouse), from a 3 month old Mdr2KO mice (n = 1 mouse) or a mouse treated with TAZ+FPC diet (n = 1 mouse). b, visualization and quantification of myHSC and cyHSC populations in HSC from healthy (n = 4 individuals) and cirrhotic (n = 4 individuals) human livers by snRNAseq. c, analysis of microarray data from isolated HSC shows that genetic HSC activation via Mx1-Cre-mediated Lhx2 deletion (n = 4 Lhx2fl/fl, n = 4 Lhx2del) resulted in enriched myHSC signature and decreased cyHSC signature; and that genetic inhibition induced by Lrat-Cre-induced Yap1 deletion (n = 5 Yapfl/fl, n = 4 YapΔHSC) exerted the opposite effect with enriched cyHSC and decreased myHSC signature in isolated HSC after treatment with 6xCCl4. d, pseudotime analysis (of HSC from TAZ+FPC and Mdr2KO) as well as cyHSC, myHSC, Col1a1 and Hgf mRNA expression in HSC using the same dataset as in a. e, In situ analysis of Hgf mRNA, visualized by RNAscope, Col1a1-GFP and Lrat-Cre-induced TdTom and subsequent quantification shows separate populations of Col1a1-GFP high Hgf low myHSC, and Col1a1-GFPlow Hgfhigh cyHSC in Mdr2 KO (n = 3 mice) and TAZ+FPC-treated livers (n = 1 mouse). f–g, analysis of Hgf and Col1a1 expression by bulk RNA-seq (Untreated (Ctrl) and 12xCCl4: n = 4 mice, FPC: n = 5 mice) (f) or scRNA-seq (n = 1 per condition) (g) of FACS-sorted HSC from untreated mice, or from CCl4-treated or TAZ-FPC-treated mice. Dot plot bar graphs are shown as mean ± SEM. In the violin plots, box plots represent the interquartile range (IQR), Q1, median and Q3, whiskers as minimum (Q1-1.5xIQR) and maximum (Q3+1.5xIQR), and outlier data as individual dots. Each data point represents one cell (a,b,d) individual (f and Mdr2KO model in e) or quantification of one field of the same liver (TAZ+FPC model in e). Scale bar 50 µm. Statistics: a: P-values, coefficient of determination (R2) and statistical significance (P value) were determined using Pearson’s. Data in d were analysed by two-tailed Mann-Whitney test. Data in b, d and g were analysed by two-tailed Mann-Whitney test. Data in f were analysed by two-tailed Student’s t-test. Raw data are given in Source Data
