Trypsin edta 10x

Lymphoprep™ gradient was purchased from Progen (Heidelberg, Germany). RPMI 1640 medium, fetal calf serum, trypsin 10x (25 mg/mL), trypsin-EDTA 10x (5 mg/mL trypsin and 2.2 mg/mL EDTA), and phosphate buffered saline (PBS, without Ca and Mg) were from PAA Laboratories Gmbh (Coelbe, Germany). L-Glutamine, penicillin, and streptomycin were from Invitrogen (Karlsruhe, Germany). Camptothecin was from Tocris (Eching, Germany); Triton-X 100, milk powder, and N,N,N′,N′-tetramethyl-1-,2-diaminomethane (Temed) were from Carl Roth (Karlsruhe, Germany); DMSO, lipopolysaccharide (LPS), phorbol, 12-myristate 13-acetate, CFSE, ammonium persulfate, bovine serum albumin, ethanol absolute, hydrochloric acid (37%), leupeptin hemisulfate, p-coumaric acid, pepstatin A, Ponceau S, trypan blue, Tween 20, and ionomycin were from Sigma-Aldrich (Taufkirchen, Germany).
Antibodies against p-38 (Thr180/Tyr182), p-ERK1/2 (Thr202/Tyr204), p-JNK, and p-c-Jun (Ser73) and the horseradish peroxidase- (HRP-) labelled secondary antibodies, anti-mouse and anti-rabbit, were from Cell Signaling Technology (Boston, USA); anti-human COX-2 was from R&D Systems (Wiesbaden, Germany); anti-human COX-1 and anti-human 5-LOX (mouse monoclonal, clone 33) were from Santa Cruz Biotechnology (Heidelberg, Germany); and mAb against β-actin was from Sigma-Aldrich (Taufkirchen, Germany).

Everolimus (07741 FLUKA) and lovastatin (M2147 SIGMA) were purchased from Sigma, St. Louis, MO, USA. For cell culture work, drugs were diluted in dimethyl-sulfoxide (DMSO, 10 mM stock solution; Sigma, D8418). DMSO was used at the appropriate dilution as control and found to be equivalent to the blank control up to concentrations of 0.4% DMSO (equivalent to 40 μM drug concentration) in the MTS assay and western blots. Dulbecco`s Modified Eagle medium–Nutrient Mixture F-12, 1:1 (DMEM/F12) media and penicillin/streptomycin were acquired from Gibco/Invitrogen (Karlsruhe, Germany), Trypsin-EDTA (10x) from PAA Laboratories (Cölbe, Deutschland), phosphate-buffered saline (PBS) and RPMI-Medium (with L-Glutamine, NaCO₃) were purchased from Sigma. Fetal bovine serum (FBS) and amphotericin B were received from Biochrom (Berlin, Germany).