Cells (25,000 per well) were seeded in 100 ml of complete growth medium in 96-well plates and incubated with chemotherapeutic agents or vehicle. T-ALL cells were split every 48 hr and AML cells were split every 72 hr. Cell viability was assessed by counting viable cells based on trypan blue vital dye staining (Invitrogen), according to the manufacturer’s instructions. Chemotherapeutic drugs included: asparaginase (pegaspargase, Shire, Lexington, MA), dexamethasone (Sigma-Aldrich), vincristine (Selleckchem, Houston, TX), doxorubicin (Sigma-Aldrich), 6-mercaptopurine (Abcam, Cambridge, UK), CHIR99021 (Selleckchem), rapamycin (Selleckchem), RAD001 (Selleckchem), AZD2014 (Selleckchem), thapsigargin (Sigma-Aldrich) and Wnt3A (R&D systems, Minneapolis, MN). BRD0705 and BRD3731 were synthesized as described (Wagner et al., 2018 ). Caspase 3/7 activity was assessed using the Caspase Glo 3/7 Assay (Promega, Madison, WI) according to the manufacturer’s instructions.
6 mercaptopurine
Manufactured by Abcam
6-mercaptopurine is a chemical compound used in research applications. It is a purine analog with a sulfhydryl group substituted for the hydrogen at position 6 of the purine ring. 6-mercaptopurine is commonly used in biochemical and cell biology studies.
Automatically generated - may contain errors
Lab products found in correlation
Vincristine, by Selleck Chemicals (2 mentions)
Doxorubicin, by Merck Group (2 mentions)
Dexamethasone (dex), by Merck Group (2 mentions)
Caspase glo 3 7 assay, by Promega (2 mentions)
Trypan blue vital dye staining, by Thermo Fisher Scientific (1 mentions)
Rad001, by Selleck Chemicals (1 mentions)
Rapamycin, by Selleck Chemicals (1 mentions)
Chir99021, by Selleck Chemicals (1 mentions)
Azd2014, by Selleck Chemicals (1 mentions)
Thapsigargin, by Merck Group (1 mentions)
Wnt3a, by R&D Systems (1 mentions)
Cytarabine, by Selleck Chemicals (1 mentions)
Asparaginase, by Merck Group (1 mentions)
Methotrexate, by Selleck Chemicals (1 mentions)
Apoptosis detection kit 2, by BD (1 mentions)
Nelarabine, by Merck Group (1 mentions)
Etoposide, by Merck Group (1 mentions)
2 protocols using 6 mercaptopurine
1
Cell Viability Assay for Leukemia Drugs
2
Chemotherapy-Induced T-ALL Cell Apoptosis
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T-ALL cells (10,000 or 40,000 per well) were seeded in 96-well plates and incubated with chemotherapeutic agents at the doses indicated below for 48 h. Chemotherapy doses were as follows unless otherwise indicated: asparaginase, 10 international units/ml (Sigma-Aldrich); dexamethasone, 10 µM (Sigma-Aldrich); vincristine, 1 µM (Selleckchem); doxorubicin, 1 µM (Sigma-Aldrich); etoposide, 10 µM (Sigma-Aldrich); cytarabine, 10 µM (Selleckchem); nelarabine, 10 µM (Sigma-Aldrich); 6-mercaptopurine, 10 µM (Abcam); and methotrexate, 10 µM (Selleckchem). Annexin V and propidium iodide staining were assessed using the Apoptosis Detection kit II (BD Biosciences), and caspase 3/7 activity was assessed using the Caspase Glo 3/7 Assay (Promega) according to the manufacturer’s instructions.
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