Gr 1 apc cy7

We performed flow cytometry to quantify the cellular composition of peritoneal cavity before cecal slurry challenge. Single-cell suspensions were prepared from peritoneal lavage samples collected following three days of zinc (or vehicle) supplementation, as described above. We suspended the cells in FACS buffer (PBS with 1% bovine albumin and 0.1% azide) and added 10% donkey serum (Invitrogen, Life Technologies, Grand Island, NY, USA) to prevent non-specific binding to cells. Cells were then stained with appropriate dilutions of the following Abs: CD11b Alex Fluor 450, Ly-6C PE Cy7, F4/80 APC, GR-1 APC Cy7 (Tonbo Biosciences, San Diego, CA, USA) and 7AAD (Life Technologies, Carlsbad, CA, USA). Non-viable cells were exclude based on 7AAD staining. CD11b⁺ cells were then gated for F4/80+ (macrophages) and GR-1⁺ (neutrophils). We analyzed all samples using FlowJo Software (FlowJo, LLC, Ashland, OR, USA).