Seven days after injury or sham treatment, unlabeled ovalbumin (OVA; Sigma), OVA-fluorescein isothiocyanate (FITC), or OVA-Alexa Fluor 647 was injected into the gastrocnemius muscles. After 24 or 48 h, total popliteal lymph node cells from individual mice were stained for CD11c and CD11b. The FL-2 channel remained free. (A) Representative dot plots of popliteal lymph node cells showing the gating strategy of OVA-FITC+ cells among total lymph node cells 48 h after OVA application. (B) CD11c and CD11b expression of gated OVA-FITC+ cells. (C) Absolute number of OVA-FITC+ dendritic cells (DCs) in the lymph nodes per mouse 24 h (n = 4 per group) and 48 h (n = 9 per group) after injury or sham treatment. (D) OVA-Alexa Fluor 647 alone or in combination with lipopolysaccharide (LPS) was injected, and the absolute number of CD11c+OVA-Alexa+ DCs in the lymph node was determined 48 h later (n = 4 per group). Data are presented as mean±SD. Symbols indicate statistical differences that were detected with analysis of variance (ANOVA). *, p<0.05; ***, p<0.001 sham treatment vs. injury. ##, p<0.01; ###, p<0.001.