RNA-seq analysis of sorted CD45+CD11b+Ly6G+ cells from C57BL/6 and Traj18-/- CRC samples and healthy controls
RNA-seq analysis of sorted CD45+CD11b+Ly6G+ cells from C57BL/6 and Traj18-/- CRC samples and healthy controls
AOM-DSS model: 7 weeks old mice were injected intraperitoneally with 10 mg/kg body weight Azoxymethane (AOM, Merck), dissolved in isotonic saline solution. After 7 days, mice were given 1% (w/v) dextran sodium sulfate (DSS MW 40 kD; TdB Consultancy) in their drinking water for 7 days followed by 14 days of recovery. The cycles were repeated 2 times and mice sacrificed at day 49. Neutrophils were sorted from tumoral lesions or healthy colon of 14 weeks/old female TRAJ18-/- [B6(Cg)-Traj18tm1.1Kro/J; JAX stock: 030524; RRID:IMSR_JAX:030524] and C57BL/6J [The Jackson Laboratory JAX stock: 000664; RRID:IMSR_JAX:000664] mice. 5 × 105 neutrophils (CD45+CD11b+Ly6G+) were sorted with a BD FACSaria and resuspended in RNAlater before storage at -80.c Total RNA from ~ 5 × 105 neutrophils (CD45+CD11b+Ly6G+) was isolated with the RNeasy micro kit (Qiagen) and RNA quality was checked with the Agilent 2100 Bioanalyzer Sequencing libraries were prepared by using the NEBNext® rRNA Depletion Kit v2 and the NEBNext® Ultra™ II Directional RNA Library Prep kits following manufacturer's instructions.