Antigen retrieval citrate buffer solution

Tissue material was collected from the indicated tissues, fixed with 4% paraformaldehyde and embedded in paraffin. 5 µm sections were cut, deparaffinized and treated with 1 × Antigen retrieval citrate buffer solution (Dako) for 20 min in a UPO 1330G microwave set to 40% power. Immunohistochemistry was carried out as described^{25 (link)}. The following primary antibodies were used: c-MYC (clone Y69, ab32072, Abcam), Ki67 (ab15580, Abcam), ColIV (ab6586, Abcam), Cytokeratin 14 (#905301, Biolegend) and CD3 (clone D7A6E, Cell Signaling Technology). Samples were imaged using a 20X objective on a Leica DM LB microscope with Studio-Lite 1.0 software (Biomedicum Imaging Unit, University of Helsinki). Whole slide scanning was done using 3DHistech Panoramic 250 FLASH II digital slide scanner (Genome Biology Unit, University of Helsinki).

Tissues were collected and fixed with 4% paraformaldehyde (PFA) and embedded in paraffin. In all, 3–5 µm sections were cut and the slides were deparafinized and incubated in 1X Antigen retrieval citrate buffer solution (Dako) in +70 °C overnight. Histochemical stainings were carried out using standard techniques. Images were taken with a Leica DM LB microscope and Studio-Lite 1.0 software (Biomedicum Imaging Unit, University of Helsinki). Images and slides were blinded for analysis.