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. 2004 Nov 15;200(10):1257–1266. doi: 10.1084/jem.20040921

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Copyright © 2004, The Rockefeller University Press

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Figure 3. — Effect of inflammatory cytokines on CD95 signaling in NPCs and differentiated neurons. (A) RNase protection assay for caspase 8 and CD95 mRNAs on 28-d NT2-neurons and NPCs untreated (Control) or treated with 200 U/ml TNF-α, 500 U/ml IFN-γ, 100 U/ml IL-1β, or a mixture of the three cytokines (Mix). L32 levels were used as loading control, and RNA of HeLa cells were used as reference control. (B) Immunoblot analysis of caspase 8 and CD95 expression on NT2-neurons and NPCs treated as in A. Cell lysates of HuT-78 and Jurkat cells were used as control. Detection of β-actin in the same membrane blot served as loading control. One representative experiment out of three performed with commercial and tissue-isolated embryonic NPCs is shown. (C) Percentage of apoptotic cells from NT2-neurons and NPCs exposed to anti-CD95 and to the cytokine mix as indicated. The results represent the mean of four independent experiments, using tissue-isolated embryonic and adult NPCs.