Fig. 1: HSCs regulate liver regeneration and injury.
From: Hepatic stellate cells control liver zonation, size and functions via R-spondin 3
a, Lrat-cre+TdTom+ mice expressing iDTR (iDTRhet) or not (iDTRWT) were injected with diphtheria toxin (DT). The TdTom+ area (n = 6 (iDTRWT) and n = 5 (iDTRhet)), Lrat and Ccnd1 mRNA (by qPCR, n = 8 (iDTRWT) and n = 11 (iDTRhet)) and the liver–body weight ratio (n = 8 (iDTRWT) and n = 8 (iDTRhet)) were determined 7 days later. b,c, iDTRWT and iDTRhet mice were treated with DT and, 1 week later, were subjected to 70% PHx (n = 7 per group) (b) or treatment with constitutive androstane receptor agonist (c), followed by Ki-67 IHC and quantification per high-power field (HPF) (n = 8 (iDTRWT) and n = 11 (iDTRhet)) as well as qPCR analysis of Mki67 (n = 8 (iDTRWT) and n = 10 (iDTRhet)). d, iDTRWT and iDTRhet mice were treated with DT. Then, 1 week later, the mice were subjected to treatment with a sublethal dose of APAP to determine the serum ALT and necrosis area in haematoxylin and eosin (H&E) sections (n = 6 (iDTRWT) and n = 4 (iDTRhet)), or with a lethal APAP dose to determine survival (n = 4 (iDTRWT) and n = 7 (iDTRhet)). e,f, iDTRWT and iDTRhet mice (n = 5 per group) were treated with DT and 1 week later were then treated with CCl4 (e; 0.5 mg per kg, n = 5 per group) or allyl alcohol (f; 60 mg per kg) to determine the serum ALT and necrosis area in H&E sections (n = 8 (iDTRWT) and n = 7 (iDTRhet)) or a lethal dose of allyl alcohol (f; 75 mg per kg; n = 10 (iDTRWT) and n = 12 (iDTRhet)) to determine survival. g,h, Primary mouse hepatocytes (Hep) were co-cultured with or without primary mouse HSCs in a contact-dependent (g; EdU, n = 6 per group; Mki67 mRNA, n = 4 per group) or contact-independent (h; EdU, n = 6 per group; qPCR, n = 3 (hepatocytes), n = 4 (hepatocytes + HSCs)) manner to determine proliferation based on EdU staining and qPCR analysis of Mki67 mRNA. Data are mean ± s.e.m. For a–h, each dot represents one biological replicate. Scale bars, 100 µm (a–h). P values were calculated using unpaired two-tailed t-tests (a–c, e, g and h, and d and f (middle and left)) or log-rank test (d and f (right)).
