Penicillin

Curcumin (Sigma, Germany), MTT (Sigma, Germany), Leptin ELISA kit (Labor diagnostika nord gmbh & co. kg, Germany), Fetal bovine serum (Gibco, USA), Phenol-red free RPMI 1640 with L-glutamine (Gibco, USA), T47D cells (Pasteur Institute of Iran), Sodium bicarbonate (Merck, Germany), Penicillin (SERVA, Germany), Streptomycin (Merck, Germany), Amphotericin B (Merck, Germany), TRIZOL Reagent (Invitrogen, USA), First-Strand Synthesis kit (Fermentas, USA), Syber Green-I reagent (Fermentas, USA).

α-T, H₂O₂, NADH, cytosine arabinoside were purchased in Sigma (Saint-Louis, MO, USA), K-252a, SL327, LY284002 and rottlerin were obtained from Calbiochem (San Diego, CA, USA), penicillin and streptomycin came from Serva (Heidelberg, Germany). The incubation media, Dulbecco’s modified Eagle Medium (DMEM) with l-glutamine, fetal calf blood serum were from Biolot Company (Saint-Petersburg, Russia). In the Immunoblotting section the information about various antibodies which were needed for experiments is given.

All cytokines and Abs used for ELISA and flowcytometry were procured from Becton Dickinson (Franklin Lakes, NJ), or otherwise mentioned. Growth media components were purchased from Hi-media (Mumbai, India) and Difco (Detroit, MI). Sodium acetate, lysozyme, proteinase K, RNase A, saponin, paraformaldehyde, CsA and other fine chemicals were purchased from SIGMA-ALDRICH (St. Louis, MO). CaeA was either procured from LKT Laboratories (St. Paul, MN) or purified from actinomycetes A. spitiensis. Aluminium sheets pre-coated with silica gel 60 F₂₅₄ were from Merck (Darmstadt, Germany). Fetal calf serum (FCS) was purchased from Harlan Sera Lab (Crawley Down, Great Britain). RPMI 1640, DMEM, HBSS and APO-BrdU TUNEL assay kit were bought from Invitrogen (Carlsbad, CA). L-glutamine, L-pyruvate, penicillin, concanavalin A and streptomycin were procured from Serva (Heidelberg, Germany). Thymidine-[methyl-³H] was the product of Amersham Pharmacia Biotech AB (Uppsala, Sweden).

The mouse motor-neuron-like hybrid cell line (NSC34), kindly provided by Dr. N.L. Cashman (University of British Columbia, Vancouver, CAN) [99 (link)], was maintained in DMEM medium supplemented with 5% fetal bovine serum (FBS)(F7524, Sigma-Aldrich), 1 mM L-glutamine (ECB3004D, Euroclone, Pero, Italy), and penicillin-streptomycin (penicillin, 31749.04, SERVA Electrophoresis GmbH, Heidelberg, Germany; streptomycin, 35500.01, SERVA Electrophoresis GmbH), at 37 °C in 5% CO₂. For experiments involving testosterone treatment, a dextran-treated charcoal-stripped FBS was used to selectively remove hormones [100 (link),101 (link)]. For the expression of the exogenous mutant proteins, NSC34 cells were transiently transfected with 0.7 μg of plasmid DNA using transferrin (T8150, Sigma-Aldrich) in conjunction with Lipofectamine reagent (18324012, Thermo Scientific Life Sciences Research, Waltham, MA, USA) to improve transfection efficiency.