scRNAseq analysis of lung with eliminating p16h cells [scRNA-seq Lung_DTR]

We employed the Chromium GEM-X single-cell RNA sequencing platform by 10x Genomics to investigate how the removal of p16-high-expressing cells affects the lung single-cell transcriptome of 22-month-old male mice. In this experiment, we utilized p16-CreERT2/R26-LSL-DTR mice, administering tamoxifen and diphtheria toxin at 21 months of age. Lung tissues were harvested two weeks later for analysis. Our objective was to examine how the elimination of p16-high cells influences intercellular interactions among diverse cell types in aged lungs. Twenty-one-month-old male p16-CreERT2/R26-LSL-DTR mice and control p16-CreERT2 mice received tamoxifen (TAM) injections for five consecutive days. Two weeks later, both groups were administered diphtheria toxin (DT) for another five consecutive days to eliminate p16-high-expressing cells. Following a two-week recovery period, lungs were harvested, and tissue dissociation was performed using a Liberase-based enzymatic digestion protocol. Single cells were then collected by fluorescence-activated cell sorting (FACS) for library preparation.