Additional file 2: Figure S1. of Decreased neuroinflammation correlates to higher vagus nerve activity fluctuations in near-term ovine fetuses: a case for the afferent cholinergic anti-inflammatory pathway?

C-Fos in fetal sheep brain. A. Western blot establishing the specificity of this antibody in near-term fetal sheep brainstem, cerebellum and cortex. B. Western blot: raw data of the image shown in Fig. S1A. C. c-Fos immunohistochemistry (IHC) in near-term fetal sheep and guinea pig brainstems. Top left: positive control staining. The cervical vagus nerve trunks were stimulated bilaterally (the stimulation was applied proximal to the bilateral cervical vagatomy to ensure strictly afferent signaling). Note diffuse c-Fos signal with high levels of background stain. Top right: negative control staining. Similar procedure was performed as in afferent stimulation, except the stimulation was performed distal of the vagatomy site ensuring strictly efferent signaling. Bottom left: example of a UCO group fetal sheep staining. Bottom right: Here we demonstrate the IHC approach regarding secondary antibody and visualization techniques; as primary antibody we used MBP (details in Methods). (ZIP 85110 kb)