Anti interleukin 1β il 1β

Per and PB were purchased from Sigma-Aldrich. Primary antibodies anti-interleukin-1β (IL-1β), anti-brain derived neurotrophic factor (BDNF) were purchased from Santacruz Biotechnology (Dallas, TX, USA). Species specific biotinylated secondary antibodies and streptavidin-HRP (Vectastain ABC Kit) were purchased from Vector laboratories (Burlingame, CA, USA). All other chemicals used in the present study were purchased from Sigma unless specified. Animal tissues were sent for paraffin embedding and sectioning to AML Laboratories (St. Augustine, FL, USA). Fecal samples from experimental mice groups and GW Veterans were sent to COSMOSID (Germantown, MD, USA) for whole-genome sequencing.

The western blot analysis was assessed as previously described with minor changes [25 (link),26 (link),27 (link)]. In brief, an equal volume of 20–30 μg of proteins (extracted from the ipsilateral cortex) was mixed with 2× Sample Buffer (Invitrogen). To separate the proteins, an equal volume of the proteins were run on 10% of SDS polyacrylamide gel electrophoresis and transferred to the PVDF membrane followed by blocking in 5% skim milk. The membranes were slightly washed to clear the skim milk. The primary antibody was incubated overnight at 4 °C 1:1000, anti-(D2R), anti-Glycogen synthase kinase 3 (p-GSK3-β) (Ser9), p-Akt (Ser473), anti-Glial fibrillary acidic protein beta (Anti-GFAP), anti-ionized calcium-binding adapter molecule 1 (anti-Iba-1), anti-phospho-c-Jun N-terminal kinase (p-JNK), anti-interleukin-1β (IL-1β), anti-caspase-3, anti-poly (ADP-ribose) polymerase-1 (Anti-PARP-1), anti-Bax, anti-Bcl-2 and anti-β-actin from Santa Cruz Biotechnology. Anti-beta actin was used as a loading control. The next day, the membranes were incubated with horseradish peroxidase-conjugated secondary antibodies diluted in 1×TBST for 1–2 h as appropriate; the immunoblots were developed using an ECL chemiluminescence system, according to the manufacturer’s instructions (Amersham Pharmacia Biotech, Uppsala, Sweden).