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. 1997 Jan 21;94(2):604–609. doi: 10.1073/pnas.94.2.604

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Copyright © 1997, The National Academy of Sciences of the USA

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Btk-dependent tyrosine phosphorylation of BAP-135. 293 cells were transfected with vector alone (lane 1), Btk alone (lane 2), epitope-tagged BAP-135 alone (lane 3), epitope-tagged BAP-135 with wild-type Btk (lane 4), or epitope-tagged BAP-135 with the following Btk mutants: R28C (lane 5), E41K (lane 6), P189A (lane 7), Y223F (lane 8), W251L (lane 9), R307K (lane 10), K430E (lane 11), and Y551F (lane 12). Total cell lysates were fractionated by SDS/PAGE, and phosphotyrosine-containing proteins were detected by immunoblotting with antibody 4G10 (P-tyr, Top). The filter was subsequently probed for Btk using Ab1280 (Btk, Middle), and BAP-135 was detected by amido black staining (p135m [stain], Bottom).