5 amino 4imidazole 1 β d carboxamide ribofuranoside aicar

Uric acid (15 mg/dl, final concentration), metformin, insulin (100 nM, final concentration) and compound C (20 μM, final concentration) were purchased from Sigma (St. Louis, MO, USA). N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG, 100 μM, final concentration) and anti-IRS1 (Ser307) and anti-phospho-IRS1 (Ser307) antibodies were purchased from Invitrogen (Carlsbad, CA, USA). 5-Amino-4-imidazole-1-β-D-carboxamide ribofuranoside (AICAR, 500 μM, final concentration) was purchased from MedChemExpress (MCE LLC, USA). A rabbit anti-GAPDH antibody was purchased from Abcam (Abcam, USA). Anti-Akt (Ser473) and anti-phospho-Akt (Ser473) antibodies were purchased from Bioworld (St. Louis Park, USA). Anti-AMPK (Thr172) and anti-phospho-AMPK (Thr172) antibodies were purchased from Cell Signaling Technology (CST, Beverly, MA, USA). Cardiomyocytes were cultured in serum-free media for 24 hours and then incubated in the presence of 15 mg/dl HUA for 24 hours. Cardiomyocytes were pretreated with either metformin (0 to 20 μmol/L) or AICAR, an AMPK activator (500 μmol/L), for 60 minutes before the addition of HUA. Other cells were preincubated with compound C for 6 hours before the addition of either metformin or AICAR. Then, 2-NBDG uptake was analyzed in cardiomyocytes. were from Millipore (Billerica, MA).

Uric acid (15 mg/dl, nal concentration), metformin, insulin (100 nM, nal concentration) and compound C (20 µM, nal concentration) were purchased from Sigma (St. Louis, MO, USA). N-(7-Nitrobenz-2-oxa-1,3diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG, 100 µM, nal concentration) and anti-IRS1 (Ser307) and anti-phospho-IRS1 (Ser307) antibodies were purchased from Invitrogen (Carlsbad, CA, USA). 5-Amino-4imidazole-1-β-D-carboxamide ribofuranoside (AICAR, 500 µM, nal concentration) was purchased from MedChemExpress (MCE LLC, USA). A rabbit anti-GAPDH antibody was purchased from Abcam (Abcam, USA). Anti-Akt (Ser473) and anti-phospho-Akt (Ser473) antibodies were purchased from Bioworld (St. Louis Park, USA). Anti-AMPK (Thr172) and anti-phospho-AMPK (Thr172) antibodies were purchased from Cell Signaling Technology (CST, Beverly, MA, USA). Cardiomyocytes were cultured in serum-free media for 24 hours and then incubated in the presence of 15 mg/dl HUA for 24 hours. Cardiomyocytes were pretreated with either metformin (0 to 20 µmol/L) or AICAR, an AMPK activator (500 µmol/L), for 60 minutes before the addition of HUA. Other cells were preincubated with compound C for 6 hours before the addition of either metformin or AICAR. Then, 2-NBDG uptake was analyzed in cardiomyocytes.
were from Millipore (Billerica, MA).