Single-cell RNA sequencing of Alkbh5-deficient AT2 cells Single-cell RNA sequencing of Alkbh5-deficient and control AT2 cells after bleomycin injuryafter bleomycin injury

After tamoxifen induction, mice received intratracheal bleomycin to induce lung injury. Fourteen days after injury, lungs were harvested and processed into single-cell suspensions.

GFP⁺ AT2 cells (reporting Sftpc-driven Cre activity) were isolated by fluorescence-activated cell sorting (FACS). To ensure optimal cell loading for the 10x Genomics Chromium platform, 40% of sorted GFP⁺ AT2 cells were mixed with 60% unsorted lung cells before encapsulation for droplet-based single-cell RNA sequencing. Libraries were prepared using the 10x Genomics Chromium Single Cell 3’ v3.1 protocol and sequenced on an Illumina platform.