Three ALK+ ALCL cell lines were used in this study: Karpas 299 (a gift of Dr. M. Kadin, Beth Israel-Deaconess Medical Center, Boston, MA, USA), SR-786, and SUP-M2 (both from ATCC, Rockville, MD, USA). The T-cell acute lymphoblastic leukemia (T-ALL) cell line Jurkat was used as a negative control because of the absence of AP-1 activity. 293T is a cell line derived from human embryonic kidney (HEK) cells (ATCC). All cell lines were grown in Roswell Park Memorial Institute (RPMI)-1640 medium (Life Technologies, Grand Island, NY, USA) as described previously12 (link), supplemented with 10% fetal calf serum, and incubated at 37°C in a humidified atmosphere containing 5% CO2.
The JUNB expression vector pcDNA3.1/V5-His Topo was a gift from Dr. Anupam Agarwal (University of Alabama at Birmingham, Birmingham, AL, USA). The AKT1 luciferase promoter (−4293/+1888/Luc) was a gift from Dr. J. Q. Cheng (University of South Florida College of Medicine, Tampa, FL, USA). STAT 3 inhibitory compound Stattic (Sigma, St. Louis, MO, USA), a small-molecule inhibitor of STAT3 activation and dimerization, and the specific MEK1/2 inhibitor U0126 (Cell Signaling Technology, Beverly, MA, USA) were used.
The JUNB expression vector pcDNA3.1/V5-His Topo was a gift from Dr. Anupam Agarwal (University of Alabama at Birmingham, Birmingham, AL, USA). The AKT1 luciferase promoter (−4293/+1888/Luc) was a gift from Dr. J. Q. Cheng (University of South Florida College of Medicine, Tampa, FL, USA). STAT 3 inhibitory compound Stattic (Sigma, St. Louis, MO, USA), a small-molecule inhibitor of STAT3 activation and dimerization, and the specific MEK1/2 inhibitor U0126 (Cell Signaling Technology, Beverly, MA, USA) were used.