NextSeq 500 sequencing; Comparison of CD8+ T-cell sub-types from peripheral lymph nodes and spleen from Balb/c or C57Bl/6J SPF mice mice by transcription profiling.
Comparison of CD8+ T-cell sub-types from peripheral lymph nodes and spleen from Balb/c or C57Bl/6J SPF mice mice by transcription profiling.
Young Balb/c or C57Bl/6J SPF mice (10-12 weeks) were irradiated with 4 Gy and sacrificed 16 days after irradiation. Lymphocytes from peripheral lymph nodes and spleen were stained with TCRβ, CD8, CD44, CD49d, CD122 and a viability dye and sorted by BD Influx to Trizol LS.  Each sample consisted of a pool of lymphocytes originating from 2-5 mice. Alternatively, young (7-12 weeks) and aged (12-19 months) Balb/c or C57Bl/6J mice were sacrificed and their lymphocytes were sorted following the same protocol.  RNA was isolated using Trizol (Thermo Fisher Scientific) followed by in-column DNAse treatment using RNA clean&concentrator kit (Zymo Research) Libraries were prepared with a SMARTer® Stranded Total RNA-Seq – Pico Input Mammalian library preparation kit v2 (Takara).