Immunofluorescent Analysis of Mouse Lung Tissue

We followed the methods of Sun et al. [17 (link)]. Four percent paraformaldehyde-fixed mouse lung tissues were cut into 4 μm thick sections. After dehydration, the sections were collected on Superfrost Plus glass slides. Sections were rinsed with PBS and permeabilized with a 1% Triton solution for 5 min. The cells were then blocked with 1% BSA for 1 h. Primary antibodies against α-SMA (Abcam, Ab240654, 1:200), SENP1 (Abcam, Ab236094, 1:200), and Sca-1 (Abcam, Ab51317, 1:200) were incubated with the sections overnight at 4 °C. Secondary antibodies were incubated with the samples for 1 h at room temperature. The sections were mounted with Fluorescent Mounting Media with DAPI and observed under a Leica SP8 confocal laser scanning microscope at magnifications of × 200.