Anti f4 80 pe cy7 clone bm8

Pancreatic leukocytes and splenocytes were stained with the following Abs: anti-CD45 APC (clone 30-F11, Biolegend, San Diego, CA, USA), anti-CD11b APC/Cy7 (clone M1/70, Biolegend, San Diego, CA, USA), anti-Ly6G FITC (clone RB6-8C5, eBioscience, CA, USA), anti-F4/80 PE/Cy7 (clone BM8, Biolegend, San Diego, CA, USA) and anti-α7nAChR PE (clone 319, Santa Cruz). Cells were suspended in Pharmingen Stain Buffer and analyzed using a ACEA NovoCyte flow cytometer with NoVo Express software (ACEA Biosciences, San Diego, CA, USA).

Pancreatic acinar cells and leukocytes were obtained using a collagenase digestion method described previously for flow cytometry analysis (Wu et al., 2018 (link)). The pancreas was removed, minced into small fragments, and digested in a DMEM medium supplemented with 2% fetal bovine serum and collagenase IV (BioFroxx, Germany, at a concentration of 2 mg/ml). Samples were incubated under agitation for 15 min at 37°C and vortexed at a low speed for 20 s before passing through a 70-mm filter. Pancreas homogenates were subjected to centrifugation to obtain pancreas acinar cells and leukocytes.
Pancreatic leukocytes were stained with the following Abs: anti-CD11b APC/Cy7 (clone M1/70, Biolegend, San Diego, CA, United States), anti-Ly6G FITC (clone RB6-8C5, eBioscience, CA, United States), anti-F4/80 PE/Cy7 (clone BM8, Biolegend, San Diego, CA, United States), and anti-α7nAChR PE (clone 319, Santa Cruz). Cells were suspended in a Pharmingen Stain Buffer and analyzed using a ACEA NovoCyte flow cytometer with NoVo Express software (ACEA Biosciences, San Diego, CA, United States).