Elisa plate reader

Cells were seeded into 96-well plates at a density of 1×10³ cells/well. Cell proliferation was assessed using the CCK-8 (Beyotime Institute of Biotechnology, Shanghai, China) assay. After treatment with GINS2 siRNA, cells were grown for 12, 24, 48, and 72 h, and 10 μL CCK-8 was added to each well and incubated for 1 h. Then, the optical density (OD) was measured at wavelength of 450 nm using an enzyme-linked immunosorbent assay (ELISA) plate reader (Promega, Madison, WI, USA). Control cells were treated with cultured media containing 0.15% (v/v) dimethyl sulfoxide (DMSO). At least three independent experiments were carried out.

Tumor necrosis factor-α (TNF-α), Nuclear factor-kappa B (NF-κB), interleukin-1β (IL-1β), and interleukin-6 (IL-6) levels in the hepatic and renal homogenates tissues were measured by commercial ELISA kits (Invitrogen, Waltham, MA, USA) following the manufacturer’s instruction. Optical densities (OD) obtained were read by ELISA Plate Reader (Glomax Multi detection system, Promega, Milan, Italy), the concentrations were expressed in nanograms per grams of tissue (ng/g tissue), and the measurements were conducted in triplicate.