A24920

Tissues were fixed in 4% PFA overnight and paraffin-processed. We cut 5-µm sections from paraffin-embedded blocks for H&E staining and immunohistochemistry. Adherent cells cultured in two-well chamber slides were stained by immunocytochemistry.
The following antibodies were used for immunofluorescence at the indicated concentrations: DDR1 (1:50; Santa Cruz Biotechnology, sc-532), E-cadherin (1:100; BD Biosciences, 610181), K8 (1:50; Developmental Studies Hybridoma Bank, TROMA-I), K14 (1:5000 [Convance, PRB-155P] and 1:100 [Santa Cruz Biotechnology, sc-17104]), α-SMA Cy3 conjugate (1:250; Sigma, C6198), vimentin (1:200; Sigma, V5255), DDR2 (1:50; LifeSpan BioSciences, LS-C164363), phH3 (1:100; Cell Signaling, 9701), HIF1α (1:50; Novus Biologicals, NB100-479), pMLC2 (1:100; Cell Signaling Technology, 3671), goat anti-mouse IgM μ chain Cy3 conjugate (1:200; Jackson ImmunoResearch, 115-166-075), and Alexa 488 anti-rat, Alexa 488 anti-rabbit, Alexa 488 anti-mouse, Alexa 568 anti-rabbit, and Alexa 647 anti-goat secondary antibodies (1:500; Molecular Probes, A11006, A24922, A24920, A21069, and A21447). Nuclei were stained with DAPI (Vector Laboratories, H-1200). Confocal or fluorescence microscopy was performed on a Nikon C1si confocal microscope or a Keyence BZ-X700 fluorescence microscope.