Frozen liver sections (7 µm) were stained with anti-PanCK and anti-CD45 (BD Pharmingen, Cat. 550539). Anti-EpCAM (Sapphire Bioscience, Waterloo, NSW, Australia, Cat. ab71916), anti-E-cadherin (Genesearch, Arundel, QLD, Australia, Cat. 3195S), anti-CK19 (TROMA III, a gift from Rolf Kemler at the Max-Planck Institute), anti-vimentin (BioScientific, Sydney, NSW, Australia, Cat. MAB2105) and anti-HNF4α (hepatocyte nuclear factor 4 alpha; Thermo Fisher Scientific, Scoresby, VIC, Cat. SCZSC-6556) antibodies were used to stain cultured LPCs. Samples were fixed in acetone-methanol (v/v, 1:1), washed in PBS, blocked in 5% bovine serum albumin (BSA) in PBS for 30 min and incubated with primary antibody (1:200 in 1% BSA/PBS) overnight at 4°C. Samples were washed and incubated with either Alexa-Fluor-488 goat anti-rabbit IgG, 594 goat-anti-rat IgG or 488 rabbit anti-goat IgG secondary antibody (Life Technologies, Cat. A11008, A11007 and A11078, respectively; 1:200 in 1% BSA/PBS) for 1 h at RT. Samples were stained with DAPI, washed and mounted with Gelvatol medium as described previously (Finch et al., 2015 (link)). Fluorescence was imaged with an IX2-ILL100 Inverted Microscope (Olympus, Macquarie Park, Australia).
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