Figure 3.

Tr14 enhances macrophage efferocytotic actions in murine peritonitis. (A–D) Self-resolving inflammation was initiated by injection of zymosan (0.1 mg/mouse, i.p.) into mice. Prior to zymosan injection, Tr14 (1.5 mL/kg or 3 mL/kg) or vehicle (veh., NaCl 0.9%) were administered i.p. once daily for six days. Peritoneal exudates were collected 4, 8 and 24 h post-zymosan injection (n = 8), and from naïve untreated mice representing time point 0 h (n = 6). (A) Infiltrated PMNs (Ly6G⁺, F4/80⁻) were stained with annexin-V and DAPI to determine non-apoptotic PMNs (Ly6G⁺, F4/80⁻, annexin-V⁻, DAPI⁻, left) and early and late apoptotic PMNs (Ly6G⁺, F4/80⁻, annexin-V⁺, DAPI^−/+, right) in the peritoneal lavages, shown as single values and mean + SEM at indicated time points. (B) Numbers of efferocytotic macrophages (F4/80⁺, Ly6G⁺) in the peritoneal lavages, given as single values mean + SEM at indicated time points. * p < 0.05, versus vehicle; two-way ANOVA with Dunnett´s multiple comparisons test. (C) Ratio of efferocytotic macrophages versus non-apoptotic PMN cell counts 4 h post-zymosan injection. Results are shown as mean + SEM. (D) Mean fluorescent intensity (MFI) of Ly6G⁺ PMNs in efferocytotic macrophages (F4/80⁺) after 4 h, measured by flow cytometry, given as % increase versus vehicle. Results are shown as mean + SEM.