Figure 3.

Variation in immune cell content in Junbo mouse bulla fluid. (a) Gating strategy applied to differentiate live cells (sytox green‐ve), neutrophils (CD11b + veLy6G + ve), macrophages (CD11b + veF4/80 + veLy6G‐veLy6C‐ve), monocytes (CD11b + veLy6C + Lvey6G‐veF4/80 ± ve), and lymphocytes (CD11b‐veCD5 + ve) present in Junbo mouse bulla fluids. The volume of ear fluids was adjusted with FACS buffer such that constant cell numbers were stained for flow cytometry and 50,000 live single cell events were counted. Neutrophil (b), macrophage (c), monocyte (d), and lymphocyte (e) cell counts were calculated from total cell count and flow‐cytometry percentages across different grades of ear fluid from noninoculated (empty bars) and inoculated (filled bars) Junbo mice. NTHi−ve–no NTHi, NTHi+ − 1 to 10² CFU/μl, NTHi++ − 10² to 10³ CFU/μl, NTHi+++ − 10³ to 10⁴ CFU/μl. n = 12, *P < 0.05,**P < 0.01