GFP-positive leukemia cells were isolated from the bone marrow of MLL-AF9 leukemia mice. The cells were maintained in Iscove’s modified Dulbecco’s medium supplemented with 10% FBS, SCF (20 ng/ml), mIL-3 (10 ng/ml; TONBO), mIL-6 (10 ng/ml; TONBO), and granulocyte-macrophage colony-stimulating factor (10 ng/ml; TONBO). The cells were plated into 96-well plates (2.0 × 104 cells per well) in the presence of GO289, incubated at 37°C for 2 days, and counted with FACSAria.

LSK hematopoietic progenitor cells as a normal counterpart were maintained in StemSpan medium (STEMCELL Technologies) supplemented with SCF (100 ng/ml) and TPO (100 ng/ml). The cells were plated into 96-well plates (1.0 × 104 cells per well) in the presence of GO289, incubated at 37°C for 2 days, and counted with FACSAria.

The human myeloid leukemia cell line, OCI-AML3, was maintained in MEM alpha medium supplemented with 20% FBS. The cells were plated into 96-well plates (1.0 × 104 cells per well) in the presence of GO289, incubated at 37°C for 4 days, and analyzed with a Cell Counting Kit-8 (Dojindo).

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



Q&A
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.