In vivo inductions of c-Fos and TNF-α in animals given LPS and AS without anesthesia.

Sixty minutes after LPS injection, MAC or EAC was given at ST36 for 30 min. Animals were sacrificed 90 min after AS for analyses of the changes in both c-Fos protein signals in the transverse sections through the caudal portion of the brainstem (A) and the TNF-α mRNA and protein in the spleen (B and C, respectively). Quantitations of both the band intensity of splenic TNF-α mRNA relative to the actin control in (B) and the c-Fos and the TNF-α signals in (A) and (C) are shown in bar graphs (mean ± SEM, n = 4 independent experiments). *p<0.05 and **p<0.01 (one-way ANOVA). Arrows in (A) and (B) denote the approximate NTS boundary between two hemispheres. cc: central canal, WP: white pulp, RP: red pulp. CTL: non-treated control. The scale bars in (A) and (C) are 100 μm.