Mbs263618

In preparation for assays, frozen brain tissue was homogenized in lysis buffer (150 mM NaCl, 10 mM Tris solution, 1% Triton X-100, pH 7.4), including protease inhibitor (Sigma-Aldrich, St. Louis, MO, USA), subsequently centrifuged at 13,000 rpm at 4°C for ten min, and the supernatant was collected. Then, the total protein content of the brain homogenate was analyzed by a previously reported protein assay [30 (link)]. ELISA levels of IL-6, inducible nitric oxide synthase (iNOS), and tumor necrosis factor-alpha (TNF-α) (MyBioSource; MBS355410, MBS263618, and MBS2507393, respectively) were assessed using commercial rat-specific ELISA kits following the manufacturer's guidelines.

In ice-cold phosphate-buffered saline, striata were homogenized to yield 10% homogenates and the content of each parameter in the striatum was determined using the matching rat-specific commercial kits according to the manufacturer’s instructions. TNF-α, BAX, and BCL2 were assessed using Cusabio (Wuhan, China) ELISA kits (Cat. No. CSB-E11987r, CSB-EL002573RA and CSB-E08854r, respectively). Moreover, MyBiosource (San Diego, CA, USA) ELISA kits were used for determination of glutamate, GABA, IL-1β, NQO1, HO-1, COX-2, caspase-3 and iNOS (Cat. No. MBS756400, MBS045103, MBS825017, MBS7606601, MBS 764989, MBS 266603, MBS7244630 and MBS 263618, respectively).

NO assay kit (ab65328, Abcam Co., Waltham, MA, USA) analyzed the content of NO in brain tissue homogenate, following the manufacturer’s instructions. The activities of TNOS (MBS723386), iNOS (MBS263618), and constitutive/endothelial nitric oxide synthase (MBS160509) were measured using kits obtained from MyBioSource consistent with the manufacturer’s instructions.