Figure 4.

Edema toxin drives significant edema and inflammatory cytokine production. (A) Mice received subcutaneous footpad injection of Vehicle (PBS) (n=6) or ET (2 µg PA + 2 µg EF) (n=6). Paw thicknesses were measured at the indicated time points. (B, C) Mice received retro-orbital injection of Evans Blue (25 mg/kg) 30 min prior to subcutaneous footpad injection of Vehicle (PBS) (n=4) or ET (2 µg PA + 2 µg EF) (n=4). (B) Representative images of ipsilateral footpads at 5 hpi. (C) The concentration of Evans Blue dye in clarified tissue lysate of the footpad at 5 hpi. (D) Mice received subcutaneous footpad injection of Vehicle (PBS) or ET (2 µg PA + 2 µg EF). Infiltration of CD45⁺Ly6G⁺ neutrophils in the footpad was measured at 5 or 24 hpi by flow cytometry. Data is from tissue combined from 3 different animals. (E) Mice received subcutaneous footpad injection of Vehicle (PBS) or ET (2 µg PA + 2 µg EF). Multiplex cytokine analysis was performed in clarified lysate of the footpad harvested at 5 hpi (n=5). Statistical analysis: (A) Two-way ANOVA with Sidak’s post-test. *p < 0.05, **p < 0.01, ****p < 0.0001. (C) Unpaired t-test. *p < 0.05. (E) Mann-Whitney test with the two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli, 5% FDR. Discoveries are marked by asterisks.