Bms 536924

Sodium Azide (20 mM, Sigma-Aldrich), Sodium Oxamate (50 mM, Sigma-Aldrich), FX11 (10 μM, Calbiochem, San Diego, CA), BMS 536924 (5 μM, Tocris, Minneapolis, MN), Afatinib (5 μM, Selleckchem, Houston, TX), Dovitinib/TKI258 (5 μM, Selleckchem), Dasatinib (5 μM, Selleckchem), PD173074 (5 μM or 20 μM, Selleckchem), Actinomycin D (5 μM, Sigma-Aldrich, St. Louis, MO), FGF2 (2 μg/mL, Cell Signaling, Danvers, MA).

The dopaminergic neuronal cell line SN4741 (RRID:CVCL_S466) derived from mouse substantia nigra [32 (link)] was grown to 70–80% confluence in culture flasks with D-MEM high-glucose containing 10% FBS, 1% penicillin/streptomycin, and 2 mM L-glutamine (Thermo Fisher Scientific, Waltham, MA, USA). Then, cells were detached with 0.25% trypsin and plated in 6-well plates at a concentration of 200,000 cells/well. For mitochondrial O₂ consumption and immunocytochemistry, wells were pre-coated with 100 μg/mL of poly-d-lysine. Cells were treated with 200 μM MPP⁺, 0.5 μM CORT [17 (link)] (Merck/MilliporeSigma, Burlington, MA, USA), and 25 ng/mL IGF-II (provided by Lilly Laboratories, Madrid, Spain) for 2.5 or 6 h [17 (link),24 (link)]. The treatments were applied in a modified Locke’s solution (137 mM NaCl, 5 mM CaCl₂, 10 mM KCl, 25 mM glucose, 10 mM Hepes, pH: 7.4) supplemented with 1% penicillin/streptomycin and 2 mM l-glutamine. BMS-536924 1 μM (Tocris Bioscience, Bristol, UK) was used to inhibit the tyrosine kinase effect of IGF-IR and InRs. Mifepristone (MIFE) 10 μM (Merck/MilliporeSigma, USA), was used to inhibit the GC effect of the CORT.