8 bromo camp

(-)-Isoproterenol hydrochloride was purchased from Sigma-Aldrich (Cat #I6504), dissolved in water/100 mM ascorbic acid to 10 mM stock, and used at 1 μM final concentration. 5′-(N-Ethylcarboxamido)adenosine (NECA) was purchased from Sigma-Aldrich (Cat #119140), solubilized in DMSO to 10 mM stock, and used at 10 μM final concentration. Forskolin was purchased from Sigma-Aldrich (Cat #F6886), dissolved in DMSO to 10 mM stock, and used at 10 μM final concentration. Shield-1 ligand for stabilization of DD-tagged proteins was purchased from Takara Bio (Cat # 632189) and added to the cell medium to 1 μM final concentration. The cell-permeable cAMP analog, 8-Bromo-cAMP, was purchased from Santa Cruz Biotechnology (Cat #sc-201564), dissolved in DMSO and used at 1 mM final concentration within 1 month of re-suspension. The CREB inhibitor compound, 666–15, was purchased from Tocris Bioscience (Cat #5661), resuspended in DMSO and used at 100 nM final. At doses higher than 100 nM, 666–15 was toxic in our cell line. H89 dihydrochloride hydrate was purchased from Sigma-Aldrich (Cat #B1427), resuspended in DMSO to 10 mM stock, and used at 10 μM final concentration. D-luciferin sodium salt (Cat #LUCNA) and coelenterazine (Cat #CZ) were purchased from GoldBio and resuspended to 100 mM in 10 mM Hepes, and 10 mM in ethanol, respectively, and stored protected from light.

JWH-133 and ACEA was purchased from Tocris Bioscience (Westwoods Business Park Ellisville, Missouri, USA). The CB2 antagonist AM630 was purchased from Cayman Chemical (Ann Arbor, MI, USA). PKA inhibitor IV and 8-Bromo-cAMP were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA.) Antibodies against CNR1, CNR2, phospho-AMPK alpha (Thr172), AMPK alpha, P21, phospho-p53 (Ser15), P53, phospho-mTOR (Ser2448) and mTOR were purchased from Affinity Biosciences (Cincinnati, OH, USA). Antibodies against p-LKB1, LKB1, PCNA and β-Actin were purchased from Abcam Trading Company Ltd (Pudong, Shanghai, China). Goat anti-rabbit IgG-HRP and goat anti-mouse IgG-HRP secondary antibodies were purchased from KPL (Gaithersburg, MD, USA). All the other chemicals were obtained from Sigma.