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Blood, Vol. 113, Issue 14, 3276-3286, April 2, 2009
The NAD biosynthesis inhibitor APO866 has potent antitumor activity against hematologic malignancies
Blood Nahimana et al.
113: 3276
Supplemental materials for: Nahimana et al
Files in this Data Supplement:
- Figure S1. Pan caspase inhibitor partially inhibited apoptosis within 72 h but not at 96h after APO866 cell exposure (JPG, 26.5 KB)
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ML-2 cells were exposed to APO866 (10nM) for 96 h in the presence or absence of zVAD-fmk (100 µM). Cells were stained with annexin V-FITC and 7-AAD to assess PS externalization and cell viability loss, respectively. Dead cells were identified as annexin V+ or 7AAD+.
- Figure S2. Wild type Jurkat cells (A) or Jurkat cells deficient in either caspase 8 (casp8 def, panel (B), FADD (FADD def, panel (C), or RIP (RIP def, panel D) were incubated for 96 h with increasing concentrations of APO866 (JPG, 48.4 KB)
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As control (E), the above cells were cultured with 100 ng ml−1 MegaFasL. Cell death was assessed by flow cytometry using annexin V and 7AAD double staining. The percentage of early apoptotic cells (annexin V+7AAD−) are shown as hatched white bars and that of late apoptotic cells (annexin V+7AAD+) are shown as solid black bars. Data shown is representative of three independent experiments.
- Figure S3. APO866 induces in time and dose dependent manners the depolarization of mitochondrial membrane in various hematological malignant cells (JPG, 85.6 KB)
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ML-2 (A), Namalwa (B), and Jurkat cells (C) were incubated without or with various concentrations of APO866 for 96h. Mitochondrial potential was measured using JC-1 staining red vs. green fluorescence .
- Figure S4. ML-2 (A), Namalwa (B), and Jurkat (C) cells were incubated with increasing concentrations (0–1mM) of nicotinamide/NAD in presence of 10nM of APO866 (JPG, 35.6 KB)
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Cell death was monitored as described in Fig. S2.
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