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2 protocols using 1 thioglycerol

1

Synthesis and Application of Triapine and Me2NNMe2

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Triapine and Me2NNMe2 were synthesized as previously described11 (link),49 (link). U0126 was purchased from Calbiochem, z-VAD-FMK from Enzo Life Sciences (New York, USA), 1-thioglycerol, thapsigargin, antimycin A, NAC, PD98059, trametinib and selumetinib from Selleck Chemicals (TX, USA). All other chemicals were from Sigma-Aldrich.
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2

Compound Preparation for Cell Culture

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We used 4OHT (H7904, Sigma) and doxycycline (D3447, Sigma) as described20 (link). Etoposide (E1383, Sigma) was dissolved in 50 mM DMSO, aliquoted and stored at −20 °C. Before use, Etoposide was added to fresh medium at a final concentration of 20 µM. Rapamycin (1292, Tocris) was dissolved in 100% ethanol at a 27.3 mM concentration, aliquoted and stored at −20 °C. Before use Rapamycin was added to fresh medium from an intermediate 27.3 μM solution at a final concentration of 20 nM. DMOG (D3695, Sigma) was dissolved in water at a 150 mM concentration, aliquoted and stored at −20 °C. Before use, DMOG was added to fresh medium at a final concentration of 1 mM. 1-Thioglycerol (M1753, Sigma) was stored at 4 °C. Before use, 1-Thioglycerol was added to fresh medium at a final concentration of 1 mM. Nutlin-3 (S1061, Selleckchem) was dissolved in DMSO at a 10 mM concentration, aliquoted and stored at −80 °C. Before use, Nutlin-3 was added to fresh medium at a final concentration of 10 μM.
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