Protease complete ultra tablets and phosphatase phosstop inhibitors

Fatty acid‐free bovine serum albumin (BSA) and palmitic acid were obtained from Sigma (St. Louis, MO, USA). Glycogen, amyloglucosidase, hexokinase, and glucose‐6‐phosphate dehydrogenase were obtained from Sigma (St. Louis, MO). [1‐¹⁴C] palmitic acid was from American Radiolabeled Chemicals (St. Louis) and D‐[U‐¹⁴C] glucose was from GE Healthcare (Little Chalfont, UK). Protease (cOmplete Ultra Tablets) and phosphatase (PhosStop) inhibitors were from Roche Diagnostics GmbH (Mannheim, Germany). The nonesterified fatty acids (NEFAs) kit was from Wako (Mountain View, CA). Glucose was measured by the glucose oxidase method using a OneTouch Ultra Mini Monitor. The rat glucagon and insulin ELISA kits were from R&D Systems (Minneapolis, MN) and Alpco (Salem, NH), respectively. All antibodies were purchased from Cell Signaling (Danvers, MA), except for P‐ACC and PGC‐1α which were purchased from Millipore (Billerica, MA), and the PEPCK antibody that was purchased from Abcam (Cambridge, MA).

Type II collagenase, isoproterenol, fatty acid (FA)-free bovine serum albumin (BSA), palmitic acid, and the free glycerol determination kit were purchased from Sigma (St. Louis, MO, USA). [1-¹⁴C] palmitic acid and D-[U-¹⁴C] glucose were purchased from American Radiolabeled Chemicals (St. Louis, MO, USA). Protease (cOmplete Ultra Tablets) and phosphatase (PhosSTOP) inhibitors were obtained from Roche Diagnostics GmbH (Mannheim, Germany). The β-actin (Cat # 4067), AMPK (Cat # 2532), pAMPK_Thr172 (Cat # 2535), AKT (Cat # 9272), pAKT_Ser473 (Cat # 9271), p38 (Cat # 9212), pp38 (Cat # 9211), hormone-sensitive lipase (HSL; Cat # 4107), and pHSL_Ser660 (Cat # 4126) antibodies were purchased from Cell Signalling (Danvers, MA, USA). The ACC (Cat # ab45174) and pACC_Ser79 (Cat # ab68191) antibodies were purchased from Abcam (Toronto, ON, Canada). ALY688 was provided by Allysta Pharmaceuticals, Belmont, CA, USA.