FIGURE 3.

Multiplex cytokines/chemokines assay. (A–D) Release of cytokines and chemokines by bacterial injection. Control (medium only), SDSE-167 (4.5 × 10⁶ CFU/mouse), GAS-476 (9.0 × 10⁶ CFU/mouse), or S. aureus (SA) strain N315 (1.0 × 10⁷ CFU/mouse) was injected i.p. into nondiabetic (heterozygous db/+) and diabetic (homozygous db/db) BKS.Cg-Dock7 mice. After 8 h, the mice were euthanized, followed by collection of blood. Serum samples were analyzed using a multiplex cytokine/chemokine assay [(A) G-CSF; (B) IL-6; (C) MCP-1, (D) IL-1α]. IL-4, IL-5, IL-7, or TNF-α was not detected using this method. Other data are shown in Supplementary Figure S3. Data represent the mean ± SD values from an experiment performed in quadruplicate. Asterisks indicate P values calculated by ANOVA are <0.01. (E–H) Time course of cytokine/chemokine release. After i.p. injection of SDSE-167 (1 × 10⁶ CFU/mouse), diabetic (db/db) and nondiabetic (db/+) BKS. Cg-Dock7 mice were euthanized at the indicated times (0, 2, 4, and 8 h), followed by the collection of serum [(E) G-CSF; (F) IL-6; (G) MCP-1; (H) IL-1α]. Data represent the mean ± SD values of an experiment performed in triplicate. ^∗P < 0.01, Student’s t-test.