Anti cd15

LF tissue samples were fixed in 4% paraformaldehyde (PFA), embedded in paraffin, and sectioned. After the sections were pre-treated with Target Retrieval Solution, pH 9 (Tris/EDTA buffer, pH 9; Dako Japan, Co., Ltd., Tokyo, Japan), endogenous peroxidases were blocked using periodic acid (Nichirei, Tokyo, Japan). We used the following antibodies as primary antibodies: anti-human Angptl2 antibody [19] (link), anti-vimentin, anti-CD3, anti-CD15, anti-CD20, anti-CD68 (Dako Japan), anti-S100A4 (Abcam, Cambridge, UK), and anti-human p-Smad3 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA). After treatment using EnVision + System-HRP-labeled Polymer (Dako Japan), the labeling was visualized using a Histofine 3,3′-diaminobenzidine (DAB) kit (Nichirei). For double immunofluorescent staining, anti-vimentin (Dako Japan) and anti-Angptl2 or anti-TGF-β1 (Abcam) were used as the primary antibodies, and Alexa Fluor 488-labeled anti-rabbit IgG and Alexa Fluor 594-labeled anti-mouse IgG (Life Technologies) were used as the secondary antibodies. Nuclei were counterstained with 4′, 6′-diamidino-2-phenylindole (DAPI).