Rabbit anti human alpha smooth muscle actin α sma

Sample sections were stained with the following primary antibodies: rat anti-mouse Mac2 (1:200; Cedarlane Corp., Burlington, ON, Canada), rabbit anti-mouse CD206 (1:300; Abcam, Cambridge, MA, United States), goat anti-mouse perilipin-1 (1:200; Abcam), rat anti-mouse CD31 (1:200; Invitrogen, North Ryde, NSW, Australia), and rabbit anti-human alpha-smooth muscle actin (α-SMA) (1:200; Abcam). After washing, the samples were incubated with donkey anti-rat-555 immunoglobulin G (1:200; Abcam), donkey anti-goat-594 immunoglobulin G (1:200; Abcam) and donkey anti-rabbit-488 immunoglobulin G (1:200; Abcam) secondary antibodies. Nuclei were stained with DAPI (Sigma). The samples were examined under a TCS SP2 confocal microscope (Leica Microsystems GmbH, Wetzlar, Germany). Leica LAS AF software was used for images analysis.

Paraffin-embedded and formalin-fixed samples (from 4 patients undergoing transplantation due to hepatitis B induced liver failure as showed in Additional file 1: Table S1, and 3 patients undergoing surgery due to hepatic hemangioma as healthy liver controls) were cut into 5-μm sections and were then processed for immunohistochemistry. After incubating with an antibody targeted against human CD163 (DakoCytomation, Glostrup, Denmark), adjacent sections were stained with either diaminobenzidine or 3-amino-9-ethylcarbazole using the Envision System (DakoCytomation, Glostrup, Denmark). For immunofluorescence analysis, tissues were stained using polyclonal mouse anti–human CD163 (DakoCytomation, Glostrup, Denmark) and rabbit anti–human alpha-smooth muscle actin (α-SMA, Abcam, Cambridge, MA, USA) or polyclonal mouse anti–human cyclooxygenase (COX)2 (Abcam, Cambridge, MA, USA) and rabbit anti–human fibroblast activation protein (FAP, Abcam, Cambridge, MA, USA), followed by Alexa Fluor 488– or 568–conjugated goat anti–mouse IgG and Alexa Fluor 568– or 488–conjugated goat anti–rabbit IgG (Invitrogen, Grand Island, NY, USA). Positive cells were quantified using ImagePro Plus software (Media Cybernetics) and detected using confocal microscopy (Leica, Germany).