Lymphopenia-induced T cell exhaustion in STING gain-of-function mouse model
Lymphopenia-induced T cell exhaustion in STING gain-of-function mouse model
Mouse splenocytes from STING GOF mice (n=5) and WT littermate controls (n=5) were stained as described above with anti-CD3e FITC (clone 145–2 C11, BD Biosciences), anti-CD4 AF700 (RM4-5, BD Pharmingen), anti-CD8a PE (53-6 .7, BD Pharmingen) and DAPI (1 μM/mL, Sigma) before positive sorting of living CD3+CD4+ or CD3+CD8+ T cells using a BD FACSAria™ Fusion flow cytometer (IGBMC Flow Cytometry Facility, Strasbourg, France). Isolated cells were at least 95% pure.  Total RNA was extracted using RNeasy® Plus Micro Kit (Qiagen) according to the manufacturer's instructions. Libraries were prepared from 10 ng RNA using SMARTer Stranded Total RNA-Seq Kit, Pico Input Mammalian (Takara) following the manufacturer's instructions. Briefly, random primers were used for first-strand synthesis, and ribosomal cDNA was cleaved by ZapR V.2 in the presence of mammalian R-probes V.2.