Ket7009

Levels of inflammatory cytokines were determined in the mouse tissues. Tissue samples were analyzed using the ELISA kits for IL-6 (KET7009; Abbkine, Wuhan, China) and TNF-α (ADI-900- 047; Enzo, Madison Avenue, New York, NY, USA), according to the manufacturer’s instructions. Standard solutions were prepared using the reagents provided in the respective kits. The plates were read at 450 nm using an Emax Plus microplate reader (Molecular Devices, San Jose, CA, USA). Proteins were quantified using mean values of duplicate samples.

Enzyme-linked immunosorbent assays (ELISAs) were used to quantify the levels of Aβ_1–42, IL-1β, IL-6, and TNF-α. To create the total lysate, the cortical tissues of the mice were weighed and homogenized in RIPA buffer containing a cocktail of protease inhibitors (Roche Science, Mannheim, Germany) and a cocktail of phosphatase inhibitors (Sigma-Aldrich, St. Louis, MO, USA). Brain tissue homogenates were analyzed using the following ELISA kits: Aβ_1–42 (KHB3544, Invitrogen, Waltham, MA, USA), IL-1β (KET6013; Abbkine, Wuhan, China), IL-6 (KET7009; Abbkine, Wuhan, China), and TNF-α (KET7015; Abbkine), according to the manufacturer’s instructions. All the experiments were analyzed in duplicate, and the mean value of the duplicate samples was calculated in all the assays. Quantification of Aβ_1–42 and cytokine levels was performed using a VICTOR X4 Multimode Plate Reader (PerkinElmer, Waltham, MA, USA).