Four h and 1 h prior to antibiotic-killed GBS challenge, C57BL/6 mice (n=3/group for A, C, and E; n=2/group for B, D, and F) were injected i.p. with vehicle, Gö6976, or Gö6983. These mice were challenged with PBS or antibiotic-killed GBS (2 × 108 cfu/mouse) i.p. Two hr later, blood and spleens were harvested. (A, B) Whole spleen lysates were prepared and phosphorylation status or expression levels of the indicated proteins were detected by Western blot assay. Actin was used as a loading control. (C, D) Total RNA was isolated from spleen and mRNA levels of the indicated cytokines and chemokines were analyzed by RT-PCR. GAPDH was used as a loading control. (E, F) Levels of the selected cytokines in serum from each mouse were analyzed by ELISA. Data represent the mean serum cytokine concentration (pg/ml) ± SD of 3 different mice. *, p < 0.05. A, C, and E are representative results from two separate experiments and B, D, and F were done once.