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2 protocols using sq22536

1

Molecular Signaling Pathway Assay

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Primary antibodies to β-actin (SC-47778, 1:1000), CaMKK-ß (SC-50341, 1:1000), and AMPKα1/2 (SC-25792, 1:1000) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Primary antibodies to p-CaMKKβ (12818S, 1:500), mTOR (2983S, 1:1000), S6 ribosomal protein (2217S, 1:1000), and p-S6 ribosomal protein (2211S, 1:1000) were purchased from Cell Signaling (Danvers, MA). Anti-p-AMPKα1/2 (11183, 1:1000) and anti-p16 (41296, 1:1000) antibodies were from Signalway (College Park, MD, USA). Anti-LC3B (NB100-2220, 1:500) and anti-p21 (NBP2-29463, 1:500) antibodies were purchased from Novus (St. Louis, MO, USA). Anti-p-mTOR (ab63552) and anti-Ki-67 (ab15580) antibodies were purchased from Abcam (Cambridge, UK). Anti-mouse (31430), anti-rabbit (31460), anti-goat (31402), and Alexa Fluor 488 (A11008) immunoglobin G secondary antibodies (1:5000) were purchased from Invitrogen (Carlsbad, CA, USA). The anti-OR2H2 primary antibody (1:1000) was purchased from Antikoerper-online.de (Aachen, Germany). L-Cis diltiazem was from Abcam (Cambridge, UK). SQ22536 and thapsigargin were purchased from Enzo Life Science (Farmingdale, NY, USA) and U73122 was from Sigma-Aldrich (St. Louis, MO, USA). Aldehyde 13-13 was obtained from Henkel (Düsseldorf, Germany). A769662, an AMPK agonist, was purchased from Cayman (Ann Arbor, MI, USA). Rapamycin was obtained from LC Laboratories (Woburn, MA, USA).
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2

Purinergic signaling modulators' effects

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NECA and withaferin A were purchased from Biomol (Hamburg, Germany). MRS1754, SCH58261, BAY60-6583 and Lucifer Yellow (LY) were purchased from Sigma-Aldrich (Taufkirchen, Germany). CGS21680 was from Merck Millipore (Darmstadt, Germany). SQ22536 was from Enzo Life Sciences (Lörrach, Germany). Adenosine-3',5'-cyclic monophosphorothioate (Rp-cAMPS) was from Biolog Life Science Institute (Bremen, Germany). NECA, withaferin A, MRS1754, SCH58261, CGS21680, and BAY60-6583 were dissolved in DMSO while Rp-cAMPS and SQ22536 were dissolved in water. All inhibitors were preincubated for 30 min prior to addition of NECA. The vehicles DMSO or water were added to control cells in all experiments at maximal concentrations of 0.1% or 0.3%, respectively.
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