Figure 2.

Variation in viable cell content in Junbo mouse bulla fluid. (a) Gating strategy applied to differentiate live (Annexin V–ve, PI‐ve), early apoptotic (Annexin V + ve, PI‐ve), late apoptotic (Annexin V + ve, PI+ve), and necrotic (Annexin V‐ve, PI+ve) cell content in Junbo mouse bulla fluids. The volume of ear fluids from different grades was adjusted by FACS buffer, such that constant cell numbers were stained for flow cytometry and 50,000 single cell events were counted. Early apoptotic (b), Late apoptotic (c), Live (d) and Necrotic (e) cell count calculated from total cell count and flow‐cytometry percentages across different grades of ear fluid from non‐inoculated (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 = 13, *P < 0.05,**P < 0.01