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Blood, Vol. 109, Issue 4, 1442-1450, February 15, 2007
Caspase-8 prevents sustained activation of NF- B in monocytes undergoing macrophagic differentiation
Blood Rébé et al.
109: 1442
Supplemental materials for: Rébé et al, Vol 109, Issue 4, 1442-1450
Flow cytometry analysis. For death receptor expression at the cell surface, anti-DR4, DR5, DcR1, DcR2, anti-TRAIL (Alexis) anti-Fas, and anti-Fas-L (PharMingen) were used.41 Lipid rafts analysis. Raft fractions were collected as previously described.43 Proteins contained in all fractions were precipitated by 6% trichloroacetic acid , resuspended in 100 µL loading buffer and the equivalent of 200 µL (fractions 1-7), 40 µL (fraction 8), and 5 µL (fractions 9-11) was analyzed by immunoblotting.
Files in this Data Supplement:
- Figure S1. Death receptor expression at the cell surface and distribution in the plasma membrane (PDF, 167 KB) -
(A) Peripheral blood monocytes (Mo) were treated for indicated times (d, days) as in Figure 1 to induce their differentiation into macrophages (Mac) or dendritic cells (DC) before flow cytometry analysis of the cell surface expression of indicated proteins (full line, specific antibody; dashed line, control antibody). (B) U937 cells were left untreated (Co) or treated with 20 nM TPA for 12 hours (TPA) before cell fractionation on a sucrose gradient. Numbers indicate cell fractions and immunoblotting with an antiflotillin antibody identifies the raft fractions. The expression of indicated proteins was analyzed by Western blot in the defined fractions. Molecular weights are in kilodaltons. One representative of at least 3 independent experiments is shown.
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