Facs acuri c6 flow cytometer

Peripheral blood was obtained by intra-cardiac puncture and collected in EDTA microtainers (BD Biosciences, CA, USA). Bone marrow cells were harvested by flushing single femur and tibia with ice cold PBS containing 2% fetal bovine serum (FBS, Thermo Fisher Scientific, MA, USA). Total and differential cell count was determined using a Hemavet hematology analyzer with veterinary software (Drew Scientific Inc., CT, USA).
Lamina propria cells were isolated and analyzed by flow cytometry as previously described ^{44 (link)} and in supplementary methods. Apoptosis of cecal tissue neutrophils was enumerated using Annexin V/7AAD-apoptosis assay kit (BD biosciences, CA, USA) as per manufacturer's instructions. Flow cytometry of apoptosis assay was performed on a BD FACS Acuri™ C6 flow cytometer equipped with CFlow^® Plus (BD Biosciences, CA, USA) software and analyzed using FlowJo V.10 (Tree Star, OR, USA).