Fibrinolytic niche is requested for alveolar type 2 cell-mediated alveologenesis and injury repair

Ali Gibran; Runzhen Zhao; Mo Zhang; Krishan G. Jain; Jianjun Chang; Satoshi Komatsu; Xiaohui Fang; Beiyun Zhou; Jiurong Liang; Dianhua Jiang; Mistuo Ikebe; Michael A Matthay; Hong-Long Ji

The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.24.006270 doi: bioRxiv preprint Scientific), pH 6.0, for 20 min at 95°C, cooled for another 20 min at room temperature, and then washed with PBS. After blocking for 1 h with 1% BSA and 4% normal goat serum, tissue sections were incubated with following antibodies: anti-proSP-C (1:500, EMD Millipore), anti-pdpn (1:300, ThermoFisher) for mouse lung tissues and 1:100 dilution for both antibodies (Abcam) for human lung slices. After 3 times washing with PBS, secondary antibodies were applied: goat anti-rabbit IgG AF488, goat anti-hamster AF568, and goat anti-mouse AF568. Tissue sections were stained with DAPI and sealed. Images were captured using a Zeiss LSM 510 confocal microscope. Images were captured containing at least 7 -8 individual 1μm optical Z-sections. Z-stacks were obtained from at least five randomly selected areas. All images were subsequently processed with ImageJ software. AT2 cells were counted using a cell counter plug-in for ImageJ software. Cells were counted in at least 5 different areas to obtain a total above 1,000 cells and then analyzed statistically.