Anti cytokeratin 7

Immunohistochemistry was performed on a Leica Bond system using the standard protocol F30. The sections were pre-treated using heat mediated antigen retrieval with citrate-based buffer (pH 6, epitope retrieval solution 1) or EDTA based buffer (pH 9, epitope retrieval solution 2) for 20 min. The sections were then incubated with antibody for 30 min at room temperature and detected using an HRP conjugated polymer system in which DAB was used as the chromogen. The sections were counter-stained with haematoxylin and mounted with Aquatex. The following antibodies were used; anti-Pentraxin 3/PTX3 1/500 [MNB1] (Abcam 90806), anti-Cytokeratin 7 1/400 [RCK105] (Abcam 9021), anti-ITGA7 1/500 (Abcam 203254), anti-MGP 1/500 (Abcam 86233), anti-TEM1 1/400 (Abcam 67273), anti-CD31 1/800 [JC70A] (DAKO 20057487), anti-CD68 1/400 (DAKO 20058607), anti-Smooth muscle actin 1/1000 (Abcam 5694), anti-periostin 1/1500 [EPR20806] (Abcam 227049), anti-APOD 1/500 (orb155698), anti-CXCL14 1/1200 (Abcam 137541), anti-Podoplanin 1/750 [D2-40] GTX31231 (lot 821903108).

Immunofluorescence was performed, as described in detail recently without modifications^{6 (link)}. Specifically, liver tissue was fixed with paraformaldehyde, cryo-protected in sucrose, and frozen in O.C.T. (Tissue-Tek). Frozen liver sections (5 μm) were permeabilized in ice-cold methanol, then room temperature 0.1% Triton X-100, and then reacted with DAPI at 0.08 ng/mL (Invitrogen, D1306) and the antibodies indicated below. Anti-eGFP (Invitrogen, #A-11122), Anti-HNF4α (Abcam, #ab41898), Anti-HAL (Sigma Prestige Antibodies, #HPA038547), anti-Cytokeratin 7 (Abcam, #ab68459), anti-albumin (Bethyl, #A80229A), anti-KIAA0319L (AAVR, Abcam, #ab105385), anti-CD68 (Invitrogen, #14068882) antibodies were used as indicated. For Figs. 3k, and 4h–k antigen retrieval was required. This was achieved by heating the slides for ten minutes in antigen retrieval buffer (Tris-EDTA, pH 9). After immunolabelling, the images were captured and analyzed on a LSM800-Airyscan microscope using ZEN Black software.