Extended Data Fig. 11: Expression and regulation of Rspo3.

a. Rspo3 mRNA in bulk RNA-seq from mouse HSCs isolated from different fibrosis models. qHSC, quiescent HSC. b. Determination of RSPO3 protein in liver extracts from healthy mice (n = 5) and mice after treatment with 40xCCl₄ injections (n = 4) by ELISA. c. scRNA-seq data showing Col1a1 expression from mice on HF-HFD at different stages of MASLD (related to Fig. 5b). d. scRNA-seq showing Col1a1 and Hgf mRNA in quiescent, activated and deactivated mouse HSCs (related to Fig. 5c). e. Zonation of TGFB1, TGFB2 and TGFB3 (left panel) from Xu et al, Nat Genet 2024 56:953-969) and qPCR for Rspo3 in primary mouse HSCs (n = 3/group) and RSPO3 in human LX-2 HSCs (n = 4/group) after treatment with rhTGFb1, rhTGFb2 and rhTGFb3 (right panel). f. UMAP showing cell annotations in human snRNA-seq from healthy, MASLD and ALD livers (related to Fig. 5e). g. sc/snRNA-seq showing RSPO3 expression and cell annotations in human livers from the indicated dataset. h. UMAP showing quiescence marker HHIP (higher in cyHSC) and activation marker COL15A1 (high in myHSC) in human snRNA-seq (related to Fig. 5g). i. RSPO3 mRNA in endothelial cells from snRNA-seq data of healthy controls (Ctrl), MASLD and patients with alcoholic cirrhosis or alcoholic hepatitis (mean + 95% confidence interval in purple). Data are shown as mean ± s.e.m. Each dot represents one biological replicate (b,e). 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 (c,d). P values were calculated using one-way ANOVA followed by Dunnett’s multiple comparisons test (a,e), unpaired two-tailed t-tests (b) or two-way ANOVA followed by Tukey’s multiple comparison test (c,d,i).