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. Author manuscript; available in PMC: 2015 Sep 1.
Published in final edited form as: J Invest Dermatol. 2014 Aug 14;135(3):861–868. doi: 10.1038/jid.2014.364

Figure 2. c-CBL inhibition induces apoptosis in CTCL and MTX enhances this effect.

Figure 2

a. Annexin V/propidium iodide flow cytometry dot plots show marked induction of apoptosis (right two quadrants of each panel) after c-CBL knockdown. In CTCL lines (MyLa, Hut-78) which express high baseline FAS death receptor, the effect is maximal with c-CBL knockdown alone. In the other samples, treatment with MTX (which upregulates low baseline FAS death receptor expression) enhances the effect of c-CBL knockdown in CTCL lines (HH, SZ4) and SS leukemic blood samples (P1, 2). a) NS siRNA, b) MTX, c) c-CBL siRNA, d) MTX plus c-CBL siRNA. Results representative of triplicate experiments.

b. MyLa and Hut-78 CTCL lines show marked apoptosis after c-CBL siRNA knockdown regardless of whether cells are treated with anti CD3/CD28 antibodies (CD+ vs CD−) but are insensitive to antibody treatment alone. In contrast, normal CD4+ blood T-cells (N) are sensitive to antibody treatment alone as well as c-CBL knockdown. a) and c) NS siRNA, b) and d) c-CBL siRNA. Results representative of duplicate experiments.