Evaluation of the impact of genetically suppression of Nupr1 in aged vs young type 2 alveolar epithelial cells (AT2).

Our data demonstrated that Nupr1 has an age-specific function in the proliferation and differentiation of lung adenocarcinoma cells that are derived from AT2 cells. However, how Nupr1 impact AT2 cells in vivo has not been tested. Using somatic genetic approaches, we performed single-cell mRNA sequencing with primary AT2 cells with control or shRNAs targeting Nupr1. Overall design: Aged (> 2 years old) and young (12-16 weeks old) Rosa26-mTmG mice were infected by either control shRNA (Ren713) or shRNAs targeting Nupr1. Due to the Cre recombinase, cells bearing shRNAs were turned green (GFP positive). Five days post-infection, mice were exposed to hyperoxia-induced alveolar injury. Four weeks after injury (when lung injury healedin general), lungs were harvested and GFP+ epithelial cells or other cells were collected for scRNA-Seq.