The lung sections were pretreated with 1:10 FcR blocking agent (Miltenyi Biotech, Gladbach, Germany) for 10 min. They were then treated with primary antibodies (1:100 dilution) as follows: goat anti-proSP-C polyclonal antibody (sc-7706; Santa Cruz Biotech, Dallas, TX, USA), rabbit anti-p38 polyclonal antibody (original production [66 (link)]), rabbit anti-proSP-C antibody (customized production [60 (link)]; Sigma-Aldrich Japan Genosys, Ishikari, Japan), and mouse anti-phospho-p38 MAPK (pT180/pY182) (clone30, 612281; BD Biosciences, NJ, USA), followed by staining with appropriate fluorescein-conjugated secondary antibodies (1:200 dilution), and 4′,6-diamidino-2-phenylindole (DAPI) was used for nuclear staining. The stained sections were observed under a fluorescence microscope (Axio Imager A2; Zeiss, Oberkochen, Germany).
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