Fig. 3: HSC-derived RSPO3 regulates hepatocyte gene expression and liver zonation.
From: Hepatic stellate cells control liver zonation, size and functions via R-spondin 3
a, GSEA of CTNNB1-regulated genes from RNA-seq data of HSC-depleted (JEDI) versus control livers; and a heat map showing the expression (Exp) of the top 15 downregulated genes from Ctnnb1ΔHep versus Ctnnb1fl/fl livers in HSC-depleted versus control livers. b, CellPhoneDB analysis showing the top HSC–hepatocyte ligand–receptor interactions in healthy mouse liver snRNA-seq data. n = 2 livers. c, snRNA-seq analysis of Rspo3 expression in healthy mouse liver. n = 2. d, RNAscope analysis of Rspo3 colocalization with TdTom+ HSCs in Lrat-cre × TdTom livers. A representative image of two technical replicates is shown. e, RSPO3 ELISA in the supernatants from primary mouse HSCs, ECs, Kupffer cells (KCs) and hepatocytes. n = 3 per group. f, Analysis of Rspo3 expression in HSCs across mouse liver zones using 100-plex spatial transcriptomics data. g, The liver–body weight ratio and qPCR analysis of Rspo3 mRNA (n = 4 per group) in HSCs from Rspo3fl/fl (n = 8) and Rspo3ΔHSC (n = 8, 7 male, 1 female) mice. h,i, The indicated WNT-target genes determined by qPCR (h) or IHC with morphometric and zone-specific quantification (i) in Rspo3fl/fl (n = 8) and Rspo3ΔHSC (n = 8, 7 male, 1 female) livers. j, Analysis of Rspo3 expression in ECs across mouse liver zones using 100-plex spatial transcriptomics data. k, Rspo3 mRNA in isolated ECs (n = 4 per group), and the liver–body weight ratio in Rspo3fl/fl (n = 6) and Rspo3ΔEC (n = 8) mice. l,m, WNT-target genes determined by qPCR (l), and IHC analysis with morphometric and zone-specific quantification (m) in Rspo3fl/fl (n = 6) and Rspo3ΔEC (n = 8) livers. Data are mean ± s.e.m. Each dot represents one cell (c) or one biological replicate (g–i and k–m). For d, i and m, scale bars, 100 µm. For the violin plots in f and j, the box plots show the interquartile range (IQR; Q1–Q3) (box limits), the median (centre line), and the minimum (Q1 − 1.5 × IQR) and maximum (Q3 + 1.5 × IQR) values (whiskers). P values were calculated using unpaired two-tailed t-tests (e, g–i and k–m) or Wilcoxon rank-sum tests (f and j). UMAP, uniform manifold approximation and projection.
