The blood glucose and lactate concentrations were measured immediately after blood collection using an automatic glucose analyzer (Free Style, Nipro Corporation, Osaka, Japan) and lactate analyzer (Lactate Pro2; Arkray Inc. Kyoto, Japan), respectively.
We measured blood glucose levels on whole venous blood using an automatic glucose monitor (One Touch Basic, Lifescan).
Serum levels of glucose, triglycerides (TG), cholesterol, total ketone bodies, and NEFA were determined using an automatic glucose monitor (One Touch, Lifescan, Neckargemünd, Germany) or commercial kits (Sigma, Munich, Germany; RANDOX, Crumlin, Northern Ireland; WAKO, Neuss, Germany, respectively).
Plasma glucose was determined using an automatic analyzer (Cobas Fara, Roche, France).
Tail vein blood glucose was checked using an automatic glucometer (Rightest GM300; Bionime) at the indicated time.
Levels of triglycerides, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, total cholesterol and fasting plasma glucose were examined using an automatic biochemical analyser (Olympus chemistry analyser AU640, Olympus Optical Co , Ltd Tokyo, Japan) using commercial kits (Olympus Optical Co , Ltd Tokyo, Japan).
After 6 or 8 weeks of MG132 (DT group) or citrate buffer (NC and DC group) injections, all rats were sacrificed and heart blood was collected to measure BUN levels and fasting blood glucose (FBG) levels, using an automatic biochemistry analyzer.
Serum lipid and glucose levels were measured using an automatic analyzer (AU2700; OLYMPUS, Tokyo, Japan) with serum creatinine levels determined by an enzymatic method.
Plasma and urinary glucose concentrations were measured using an automatic analyzer TBA-120FR (Toshiba Medical Systems Corp., Otawara, Japan) and Glucoroder-NX (A&T Corp., Yokohama, Japan), respectively.
Plasma glucose concentrations were measured using an automatic analyzer (YSI Stat2300, Yellow Spring Instruments, Yellow Spring, OH) and plasma insulin concentrations were determined by ELISA (Invitrogen, UK).
Arterial oxygen and carbon dioxide partial pressures and blood glucose levels were measured by using an automatic blood gas analyzer (GEM 4000; Instrumentation Laboratory GmbH, Munich, Germany).
