Anti akt and anti phospho akt ser473 antibodies

DPI and 3-bromopyruvate were purchased from Sigma-Aldrich (St. Louis, MO). CM-H2DCF-DA, DAF-FM, HEt, and 2-NBDG were purchased from Invitrogen/Molecular Probes (Carlsbad, CA). SP600125 was acquired from EMD Biosciences (Calbiochem, San Diego, CA). DPI was dissolved in dimethyl sulfoxide (DMSO) and freshly diluted in culture media before used. The final DMSO concentration was less than 0.1% (v/v). In addition, 3-bromopyruvate was dissolved in water and neutralized with NaOH immediately before use in cell culture. The rabbit polyclonal anti-p22^phox antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit polyclonal anti-Akt and anti-phospho-Akt (Ser473) antibodies as well as rabbit monoclonal anti-c-Jun and anti-phospho-c-Jun (Ser63) antibodies were purchased from Cell Signaling Technology, Inc. (Beverly, MA).

Resveratrol was purchased from SIGMA (Saint Louis, Missouri, USA). Radioactive labeled [7(N)-³H]-pregnenolone (12.6 Ci/mmol) and [1,2,6,7(N)-³H]–DHEA (63 Ci/mmol) were obtained from PerkinElmer (Waltham, MA, USA). [4-¹⁴C]-progesterone (ART-1398) was purchased from American Radiolabeled Chemicals (St. Louis, MO, USA). Antibodies against human CYP17A1 and POR were custom made by GenScript (Piscataway, NJ, USA). Anti-CYP21A2 antibody was a generous gift of Prof. Walter L. Miller (San Francisco UCSF, CA, USA). The phospho-Akt pathway was studied using anti-Akt and anti-phospho-Akt (Ser 473) antibodies from Cell Signaling Technology (Danvers, MA, USA). Antibodies against SIRT1 (Abcam, Cambridge, UK), SIRT3 and SIRT5 (Cell Signaling, Danvers, MA, USA) were kindly shared by Dr. Jean-Marc Nouffer (University of Bern, Bern, Switzerland). β-actin antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA). SIRT3/5 plasmids were obtained from the laboratory of Dr. Eric Verdin (University of California San Francisco, San Francisco, CA) via Addgene (Cambridge, MA) and provided by Prof Marion B. Sewer (UC San Diego, CA, USA). SIRT1 plasmid was obtained from Prof. Anna Biason-Lauber (University of Fribourg, Fribourg, Switzerland).