Cobas c 111 bio analyzer

The liver was extracted as described previously (13 (link)), and its triglyceride and cholesterol contents were determined using the Cobas C-111 bio-analyzer (Roche, Switzerland) and normalized to wet tissue weight.

Serum insulin and urine albumin were measured by ELISA (Millipore and Bethyl Laboratories, respectively). Serum urea, glucose, alanine transaminase (ALT), alkaline phosphatase (ALP), urine glucose and creatinine, total triglycerides, cholesterol, and high- and low-density lipoprotein (HDL and LDL, respectively) were determined using the Cobas C-111 bio-analyzer (Roche, Switzerland). Blood urea nitrogen (BUN) was calculated by serum urea levels (BUN mg/dl = Urea mM × 2.801). Blood glucose was measured with the Elite glucometer (Bayer, Pittsburgh, PA). Glucose and insulin levels were used to calculate the homeostasis model assessment-insulin resistance index (HOMA-IR) as glucose (mg/dl) × insulin (U/L) 22.5. Whole kidney proteins were extracted and the levels of the following markers (MCP1, TNFα, TGFβ, and IL18) were measured using ELISA kits (Abcam; Cat# ab100721, ab46105, ab119557, and ab216165, respectively). Liver proteins were extracted and the levels of TNFα were measured using the same ELISA kit used for the kidney.