L1cam

Cell lysates were prepared in RIPA buffer (Sigma, St Louis, MO, USA) supplemented with protease inhibitors. Protein samples were separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes. The membranes were blocked and probed with primary antibody against LOXL2 (Origene; Rockville, MD, USA), Snail (Cell Signaling; Danvers, MA, USA), pFAK and FAK (Santa Cruz, CA, USA), pSRC (Millipore, Billerica, MA, USA), SRC (Santa Cruz, CA, USA), CDH1 (BD Biosciences; Sparks, MD), L1CAM (Novus Biologicals, Littleton, CO, USA), N-cadherin (Invitrogen, CA, USA), Vimentin (Dako, Glostrup, Denmark) and β-actin (Sigma, St Louis, MO, USA). The intensity of the western blotss was quantified by densitometric scanning with ImageJ (NIH) and normalized to β-actin.

Three‐micrometer‐thick tissue sections were cut and stained with hematoxylin‐eosin or periodic acid–methenamine silver (PAM). Immunohistochemical staining was performed on FFPE tissue sections. Primary antibodies against the following antigens were applied: vimentin (V9; 1:200; Dako, Glostrup, Denmark), glial fibrillary acidic protein (GFAP) (1:5000) (15 (link)), Olig2 (1:5000) (16 (link)), cytokeratin (CAM5.2; 1:5; BD Bioscience, San Jose, CA, USA), α‐smooth muscle actin (αSMA) (1A4; 1:3200; BioMakor, Rehovot, Israel), epithelial membrane antigen (EMA) (E29; 1:100; Dako), synaptophysin (27G12; 1:200; Novocastra, Newcastle upon Tyne, UK), NeuN (A60; 1:1000; Chemicon, Temecula, CA, USA), podoplanin (D2‐40; prediluted; Nichirei, Tokyo, Japan), CD99 (12E7; 1:50; Dako), L1CAM (UJ127; 1:100; Novus Biologicals, Littleton, CO, USA), p65/RelA (D14E12; 1:400; Cell Signaling Technology, Danvers, MA, USA), BAF47/INI1 (BAF47; 1:100; BD Bioscience, San Jose, CA, USA), BRG1 (polyclonal; 1:1000; Millipore, Temecula, CA, USA), and Ki‐67 (MIB‐1; 1:100; Dako). For coloration, a commercially available biotin‐streptavidin immunoperoxidase kit (Histofine, Nichirei) and diaminobenzidine were employed.