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Blood, Vol. 110, Issue 5, 1621-1630, September 1, 2007
NPM-ALK oncogenic kinase promotes cell-cycle progression through activation of JNK/cJun signaling in anaplastic large-cell lymphoma
Blood Leventaki et al.
110: 1621
Supplemental materials for Leventaki et al, Vol. 110, Issue 5, 1621-1630
Files in this Data Supplement:
- Figure S1. NPM-ALK kinase activity is not affected by JNK activity in NPM-ALK+ ALCL cells (JPG, 16.2 KB)
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NPM-ALK+ ALCL cells were transiently transfected with 40 µg of siRNA selectively targeting JNK1 or JNK2 gene products as shown in Figure 4E. Western blot analysis was performed using whole cell lysates and an antibody specific for the phosphorylated (activated) form of ALK. Immunoblot shows that ALK phosphorylation indicating kinase activity does not change after silencing JNK1 or JNK2 genes.
- Figure S2. Apoptosis after inhibition of JNK/c-Jun signaling in NPM-ALK+ ALCL cells (JPG, 50.6 KB)
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A. In addition to cell cycle changes, treatment of NPM-ALK+ ALCL cells with a specific JNK inhibitor, SP600125, for 48 hours resulted in increased apoptosis as assessed by annexin V/propidium iodide (PI) staining and flow cytometry. The percentage of annexin V/PI+ apoptotic cells was increased from 5.53% (untreated cells) to 14.63% (10 µM SP600125), 12.19% (20 µM SP600125) and 17.04% (40 µM SP600125). B. Apoptosis also was assessed in NPM-ALK+ ALCL cells after silencing of JNK1 or JNK2 genes. SU-DHL1 cells were transiently transfected with 20 µg of JNK1 or JNK2 siRNA as shown in Figure 4E, and annexin V staining was performed. The proportion of annexin V/PI+ apoptotic cells was increased from 20% (control siRNA) to 35.64% (JNK1 siRNA) and 39.58 % (JNK2 siRNA). In contrast, no detectable changes in apoptosis were observed after silencing c-Jun.
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