MEFs were extracted from E13 embryos. Briefly, embryos were sterilized with ethanol, washed with PBS and triturated with razor blades. Samples were then incubated in DMEM (Gibco-Invitrogen, Carlsbad, CA, USA) overnight at 37 °C and 5% CO2. The next day, cultured cells were trypsinized, filtered and washed. Finally, MEFs were incubated at 37 °C and 5% CO2 and used for the corresponding experiments. For autophagy induction, MEFs were cultured during 4 h with 250 nM Torin (S2827, Selleckchem, Houston, TX, USA), 100 μM spermidine (Sigma-Aldrich), 100 μM Resveratrol (Sigma-Aldrich) in supplemented DMEM. For autophagy experiments, MEFs were cultured during 4 h in the presence of 100 nM Bafilomycin A1 (BML-CM110, Enzo Life Sciences, Farmingdale, NY, USA).
Bafilomycin a1 bml cm110
Manufactured by Enzo Life Sciences
Sourced in United States, Switzerland
Bafilomycin A1 (BML-CM110) is a laboratory reagent provided by Enzo Life Sciences. It is a specific inhibitor of vacuolar-type H+-ATPases (V-ATPases).
Automatically generated - may contain errors
2 protocols using bafilomycin a1 bml cm110
1
Isolation and Autophagy Induction in Mouse Embryonic Fibroblasts
2
Starvation and Autophagy Induction in H1299 Cells
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The human non-small cell lung cancer cell line H1299 was directly obtained from Leibniz-Institute DSMZ-German collection of Microorganisms and Cell Cultures and cultured in RPMI-1640 (Sigma-Aldrich, Buchs, Switzerland) supplemented with 10% FBS (Sigma-Aldrich, product number: F7524, Lot number: 012M3396) and antibiotics. For starvation experiments cells were seeded and the next day washed with PBS before the addition of EBSS (#E2888, Sigma-Aldrich). Cells were starved for 6 h and Bafilomycin A1 (BML-CM110, Enzo-Life Sciences, Lausen, Switzerland) was added for the last two hours at a final concentration of 200 nM.
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