Western Blot Analysis of Neuroinflammatory Markers

Tissue homogenates from the hippocampus and PFC were subjected to Western Blot (WB) analysis for the determination of protein levels. The extraction of protein and WB was performed as previously described [9 (link)–12 (link)]. The primary antibodies IL1b, IL 4, IL-18, TLR4, HMGB 1 and GAPDH were commercially obtained from Abcam, Cambridge, MA. Secondary antibodies were obtained: anti-mouse, 1:5000 dilution and anti-rabbit, 1:5000 dilution (SC-2314 and SC-2004 respectively, Santa Cruz Biotechnology, Santa Cruz, CA). The Immunoreactive bands were visualized using enhanced chemiluminescence (ECL Plus, Amersham), band intensities were quantified using ImageJ imaging software (NIH), and were normalized with GAPDH.