Extended Data Fig. 7: R-spondin3 promotes hepatocyte proliferation and rescues HSC-depleted livers.
From: Hepatic stellate cells control liver zonation, size and functions via R-spondin 3
a. qPCR (n = 4/group) for indicated genes and WST-1 assay (n = 3/group) in AML12 hepatocytes treated with the indicated concentrations of Rspo3 for 24 h (qPCR) or 48 h (WST-1 assay). b. Effect of HSC co-culture on hepatocyte proliferation in the absence or presence of Rspo3-blocking antibody rosmantuzumab or control antibody. c. Schematic diagram showing AAV8-CMV-GFP or AAV8-CMV-Rspo3 rescue experiments in HSC-depleted mice followed by 70% PHx or treatment with APAP (300 mg/kg). d. Rspo3 mRNA was determined by qPCR in non-depleted control livers and liver from HSC-depleted iDTR mice after treatment with AAV8-CMV-GFP (iDTRwt n = 4, iDTRhet n = 3) or AAV8-CMV-Rspo3 (n = 4). e. Proliferation was determined 48 h after 70% PHx by Ki-67 IHC in livers from AAV8-CMV-GFP-treated WT (n = 4) and iDTRhet (n = 3) and in AAV8-CMV-Rspo3 treated iDTRhet mice (n = 4). f. Liver injury was determined by serum ALT and necrosis quantification in H&E liver sections 24 h after APAP treatment in AAV8-CMV-GFP-treated WT (n = 8) and iDTRhet (n = 7) and in AAV8-CMV-Rspo3 treated iDTRhet mice (n = 7). nd ** P < 0.01 vs ctrl, *** P < 0.001 vs ctrl. Data are shown as mean ± s.e.m. Each dot represents one biological replicate (a,b,d-f). Scale bars 100 µm (b,e,f). P-values were calculated using unpaired two-tailed t-tests (a-d).
