Phospho akt and pan akt antibodies

For western blot analyses, we utilized phospho-Akt and pan-Akt antibodies (Cell Signaling) and UCP11-A antibody from Alpha Diagnostic International (Interchim, Montluçon, France). RNA expression was analysed as previously described16 (link).

Insulin was purchased from Sigma-Aldrich (St. Louis, MO, USA). [methyl-³H]Choline chloride and [³H]HC-3 were from Perkin-Elmer (Waltham, MA, USA). 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) was from Millipore (Etobicoke, ON, Canada). All cell culture media and reagents were from Life Technologies (Burlington, ON, Canada). EZ-link sulfo-NHS-SS-biotin and Neutravidin agarose resin were from Thermo Fisher Scientific (Ottawa, ON, Canada). AlexaFluor-555 anti-Rb Zenon labeling kits and all fluorescent secondary antibodies were from Molecular Probes, Life Technologies (Burlington, ON, Canada). Polyclonal CHT antibody was prepared as described previously [29 (link)], anti-FLAG antibody was from Sigma-Aldrich (Oakville, ON, Canada), anti-actin antibody was from Santa Cruz Biotechnology (Santa Cruz, CA, USA), phospho-Akt and pan-Akt antibodies were from Cell Signaling Technology (Danvers, MA, USA) and horseradish peroxidase-conjugated goat anti-rabbit and goat anti-mouse secondary antibodies were from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). Cell culture dishes were from VWR (Mississauga, ON, Canada) with the exception of glass-bottomed dishes supplied by MatTek Corp (Ashland, MA, USA) (0.16–0.19 mm coverslip thickness) used for TIRF imaging.