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. 2005 Jun 6;201(11):1837–1852. doi: 10.1084/jem.20042101

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Copyright © 2005, The Rockefeller University Press

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Figure 3. — Specific silencing of full-length BTK or BTK splice variants by RNA interference. BCR-ABL1 ⁺ pre–B lymphoblastic leukemia cells were transfected with a nontargeting siRNA duplex in the presence or absence of STI571 (for 12 h), as well as with a mixture of siRNAs against full-length BTK, BTK^p52, or BTK^p65. All siRNA duplices were labeled with fluorescein before transfection. Transfection was repeated after 24 h, and 10⁵ cells carrying fluorescein-labeled siRNAs (between 3–5% of all viable cells) were sorted by FACS 48 h after the first transfection (upper panel). Total RNA was isolated and subjected to semiquantitative RT-PCR analysis of BTK isoform expression (lower panel). mRNA levels for HPRT and COX6B (see Fig. 8 C) were stable under all conditions.