TertHigh Cells: Key Players in Salivary Gland Homeostasis and Regeneration after Radiation Therapy in Adult Mice

To gain insight into the mechanisms by which TertHigh populations regulate cell survival post radiation treatment, we conducted RNA-Seq analysis of sorted TertHigh and TertLow SMG cells treated with or without RT. In this experiment, 2-month-old TertCreERT2/+Rosa26LSL-tdTomato/+ mice were treated with tamoxifen, followed by RT after a 7-day interval. Subsequently, the SMGs were harvested and dissociated 36 hours after RT treatment. We isolated and separately sorted 10,000 TertHigh and TertLow cells for RNA sequencing. Overall design: After sorting TertHigh (DAPI-EpCAM+tdTomato+) and TertLow (DAPI-EpCAM+tdTomato-) cells from SMGs treated with or without RT, cells were subjected to RNA extraction and library preparation. 1ng of purified RNA was used for preparing the library according to the Dr. Picelli's protocol. Illumina dual Indices were used for library multiplexing. RNA sequencing was performed by Novogene via Illumina platforms based on mechanism of SBS (Sequencing by synthesis).