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. 2001 Sep 17;194(6):863–870. doi: 10.1084/jem.194.6.863

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© 2001 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Quantitation of TLR mRNA by real-time quantitative RT-PCR in freshly isolated monocytes (Mono), CD11c⁺ imDCs, and plasmacytoid pre-DCs (pDC), and in immature and mature DCs induced from them. Monocytes were cultured with 50 ng/ml GM-CSF and 200 U/ml IL-4 for 5 d to obtain imDC1 and were further stimulated with CD40L-transfected L cells for 24 h to obtain mDC1. CD11c⁺ imDCs were stimulated with CD40L-transfected L cells for 24 h to obtain mature CD11c⁺ DCs. Plasmacytoid pre-DCs were cultured with 10 ng/ml IL-3 for 5 d to obtain imDC2 and were further stimulated with CD40L-transfected L cells for 24 h to obtain mature DC2. The amounts of mRNA were quantitated by real-time quantitative RT-PCR, and were shown by arbitrary unit relative to the amount of ubiquitin mRNA. The data shown are representative of three experiments.