N n n n tetramethylenediamine

RPMI 1640, penicillin, streptomycin and PBS were purchased from Gibco, Lifetechnologies (Darmstadt, Germany). FCS was purchased from Biochrom (Berlin, Germany). RIPA buffer, protein inhibitors, molecular mass standards for SDS-PAGE, DMSO, TX-100, Histopaque, Sodium dodecyl sulphate (SDS), 5,6,6-tetrachloro-1,1,3,3-tetraethylbenzimidazol-carbocyanine iodide (JC-1), carbonyl cyanide 3-chlorophenylhydrazone (CCCP) and propidium iodide (PI) were obtained from Sigma-Aldrich (Munich, Germany). Tween, Sulphuric acid, acrylamide and dithiotreitol were purchased from Carl Roth GmbH, (Karlsruhe, Germany). Ammonium persulfate and N,N,N,N-tetramethylenediamine were obtained from BioRad (Munich, Germany). 3,3’,5,5’-tetramethylbenzidine was purchased from eBioscience Inc. (San Diego, USA). Following primary antibodies were used: caspase-3, poly (ADP-ribose) polymerase (PARP), claspin, survivin, bcl-2, cytochrome c (Cell Signaling Technology, Danvers, USA); p53 (Santa Cruz biotechnology, Santa Cruz, CA, USA); X-chromosome-linked IAP (XIAP) and Annexin V-APC (BD Bioscience, Heidelberg, Germany); ß-actin-peroxidase antibody (Sigma-Aldrich, Munich, Germany). The Cytotoxicity Detection Kit was purchased from Roche (Grenzach-Wyhlen, Germany).

Molecular weight standards, acrylamide, SDS, N,N′-methylene-bisacrylamide, N,N,N′,N′-tetramethylenediamine, protein assay reagents, and nitrocellulose membranes were purchased from Bio-Rad Laboratories (Hercules, CA, USA). Protein kinase C inhibitor Ro-32-0432 was from Calbiochem (La Jolla, CA, USA). Rabbit polyclonal anti-prostasin antibodies, anti-phospho-Sgk-1 (S422) and anti-total-Sgk-1 antibodies were obtained from Abcam Inc. (Cambridge, MA, USA). The anti-Sgk-1 (T256) antibody, the rabbit anti-total human Akt polyclonal antibody, as well as mouse anti-human phospho-Akt (S473) and anti-human phospho-Akt (T308) mAb were purchased from Cell Signaling (Danvers, MA, USA). Anti-beta-ENaC antibodies were purchased from ProteinTech (Rosemont, IL, USA) and anti-gamma-ENaC antibodies were from Abcam (Cambridge, MA, USA). Rabbit polyclonal anti-human ENaC-alpha antibodies were either made in-house (ENaC-alpha 60) [35 (link)] or were purchased from Novus Biologicals (Littleton, CO, USA). The Pierce supersignal substrate chemiluminescence detection kit was obtained from Pierce (Rockford, IL, USA). Monoclonal anti-β-actin antibody, fibronectin, Tween-20, and all other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).