Quantifying Apoptosis-related Proteins

Proteins were quantified by immunoblot analysis as described previously (21 (link), 24 (link)). For co-culture experiments, leukemia cells were collected by combining cells in the supernatant and after two washes with PBS. Antibodies against p-CRKL, BID, and BCL-X_L were purchased from Cell Signaling Technology, MCL-1 from Santa Cruz Biotechnology Inc., BIM and CRKL from Abcam, BAX from Sigma, and BCL-2 from Dako. Signals were detected using an Odyssey infrared imaging system and quantitated using the Odyssey software program (version 3.0; LI-COR Biotechnology). β-ACTIN (Sigma) was used as a loading control.

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Carter B.Z., Mak P.Y., Mu H., Zhou H., Mak D.H., Schober W., Leverson J.D., Zhang B., Bhatia R., Huang X., Cortes J., Kantarjian H., Konopleva M, & Andreeff M. (2016). Combined Targeting of BCL-2 and BCR-ABL Tyrosine Kinase Eradicates Chronic Myeloid Leukemia Stem Cells. Science translational medicine, 8(355), 355ra117.

Publication 2016

p-CRKL BCL-XL MCL-1 BAX BCL-2 Odyssey infrared imaging system Odyssey software program β-ACTIN

Actin Antibodies Bcl 2 Cells Co culture Crkl Immunoblot analysis Leukemia Proteins

Corresponding Organization : The University of Texas MD Anderson Cancer Center

Other organizations : AbbVie (United States), City of Hope, University of Alabama at Birmingham

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Variable analysis

independent variables

Co-culture experiments

dependent variables

Protein quantification by immunoblot analysis

control variables

Cells were washed with PBS
β-ACTIN was used as a loading control

controls

Positive control: None specified
Negative control: None specified

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