KT-1 (64) and U937 (CRL-1593.2; American Type Culture Collection) cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and antibiotics. 2fTGH cells (65) were provided by G. R. Stark (Cleveland Clinic) and were cultured in DMEM (Dulbecco’s modified Eagle’s medium) high-glucose medium supplemented with 10% FBS and antibiotics. The immortalized Ulk1/2+/+ and Ulk1/2−/− MEFs (17) were provided by C. B. Thompson (Memorial Sloan-Kettering Cancer Center) and were cultured in DMEM supplemented with 10% FBS and antibiotics. Primary Mlk3+/+ and Mlk3−/− MEFs were described previously (12) and were cultured in DMEM high-glucose medium supplemented with 10% bovine growth serum, 1× l-glutamine, 1× penicillin/streptomycin solution, and 8 μl of 2-mercaptoethanol per liter of medium. All experiments were performed using primary MEFs between passages 2 and 4. The other cell lines were frozen at low passage in liquid nitrogen and were kept in culture for no longer than eight passages. All cells were cultured at 37°C and 5% CO2. All cells were tested for mycoplasma contamination using a MycoAlert PLUS mycoplasma detection kit, following the manufacturer’s instructions (Lonza).

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.