FACS Sorting of Lung AEC2 Cells

For FACS sorting, lungs from WT or Sftpc-CreER; tdTomato⁺ mice were collected at 6 to 8 weeks of age and processed into a single-cell suspension using dispase, DNase I and collagenase I, as previously described [15 (link)]. The AEC2 population (Sftpc⁺ AEC2 cells) was isolated from the lungs of 6–8-week-old Sftpc-CreER; tdTomato⁺ mice 5 days after induction with 200 μg/g body weight tamoxifen. Krt5⁺ AEC2s and Krt5⁻ AEC2s were sorted by MoFlo SX (Beckman Coulter, Miami FL). Data were analyzed in Summit 5.2 (Beckman). For FACS analysis and sorting of mouse, the following antibodies were used: CD45 (Biolegend, 103,116, 1:100), LysoTracker (Invitrogen, L7526, 1:14000), EpCAM (Biolegend, 118,206, 1:100), Krt5 (Abcam, ab193895, 1:300), and Edu (50 mg/kg, E10187, Life Technologies). For the FACS analysis and sorting of patients, the following antibodies were used: LysoTracker (Invitrogen, #L7526, 1:14000), CD45 (Biolegend, #368,530, 1:1000), KRT5 (ABcam, #ab193895, 1:1000); EpCAM (Biolegend, #324,220, 1:1000).