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Blood, Vol. 111, Issue 7, 3793-3801, April 1, 2008
HO-1 underlies resistance of AML cells to TNF-induced apoptosis
Blood Rushworth and MacEwan
111: 3793
Supplemental materials for: Rushworth and MacEwan
Files in this Data Supplement:
- Figure S1. CD34+ hematopoietic stem cells are not resistant to TNF induced cell death (JPG, 43 KB)
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(A) CD34+ hematopoietic stem cells, primary monocytes, THP-1, HL60, were treated with 10 µM BAY 11-7082 30 min prior to treatment with the indicated concentrations of TNF for 24 h. Cell number was then assessed by MTS assay as described in the Materials and Methods section. (B) CD34+ hematopoietic stem cells, primary monocytes, THP-1 cells and HL60 cells were treated with BAY-11-7082 (10 µM) or transfected prior to treatment with TNF (10 ng/ml) for the indicated time. RNA was extracted and after reverse transcription, HO-1 mRNA expression was measured by real-time PCR. Values represent means ± SEM from three independent experiments.
- Figure S2. FLIP has a limited role in regulating cell death in response to TNF in combination with NF-κB inhibition (JPG, 78.3 KB)
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(A) THP-1 cells and HL60 cells were treated with various concentrations of BAY 11-7082 30 min prior to treatment with 10ng/ml of TNF for 24 h. Cell number was then assessed by MTS assay. (B) THP-1 cells and HL60 cells were treated with 1ug/ml of CHX prior to treatment with 10 µM BAY 11-7082, or 10 ng/ml TNF, or BAY 11-7082 plus TNF. Cell number was then assessed by MTS assay. (C). THP-1 cells were treated with 1ug/ml of CHX prior to treatment with BAY 11-7082 plus TNF. Western blot analysis was carried out using anti-FLIP, anti-Nrf2 and anti- -actin antibodies. (D) THP-1 cells and HL60 cells were treated with 1 µM of geldanamycin prior to treatment with 10 µM BAY 11-7082, or 10 ng/ml TNF, or BAY 11-7082 plus TNF. Cell number was then assessed by MTS assay. Values represent means ± SEM from three independent experiments.
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