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. 2000 Jun 12;149(6):1249–1262. doi: 10.1083/jcb.149.6.1249

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© 2000 The Rockefeller University Press

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graphic file with name JCB9907095.f1a.jpg — PYK-2 is localized at the uropod projection of NK cells adhered to fibronectin. (a) Confocal section of PYK-2 immune staining of NK cells migrating on FN (A). The same section under brightfield illumination is shown in B. Epifluorescence images of NK cells double-stained for PYK-2 (green) and moesin (red, C), and PYK-2 (green) and talin (red, D). Control cells stained with goat serum or secondary antibody alone did not show any specific staining, and preincubation of the anti-PYK-2 antiserum with its specific blocking peptide prevented staining of NK cells (not shown). (b, A) Confocal serial sections of polarized NK cells adhered to FN and double-stained for γ-tubulin (red, upper panels, γ-TUB) and PYK-2 (green, lower panels, PYK-2). Sections were taken every 0.8 μm from the substratum (0) in the z-axis. b, B shows a merged image of double-labeled cells demonstrating the colocalization of γ-tubulin and PYK-2 at the uropod of NK cells.

graphic file with name JCB9907095.f1b.jpg — PYK-2 is localized at the uropod projection of NK cells adhered to fibronectin. (a) Confocal section of PYK-2 immune staining of NK cells migrating on FN (A). The same section under brightfield illumination is shown in B. Epifluorescence images of NK cells double-stained for PYK-2 (green) and moesin (red, C), and PYK-2 (green) and talin (red, D). Control cells stained with goat serum or secondary antibody alone did not show any specific staining, and preincubation of the anti-PYK-2 antiserum with its specific blocking peptide prevented staining of NK cells (not shown). (b, A) Confocal serial sections of polarized NK cells adhered to FN and double-stained for γ-tubulin (red, upper panels, γ-TUB) and PYK-2 (green, lower panels, PYK-2). Sections were taken every 0.8 μm from the substratum (0) in the z-axis. b, B shows a merged image of double-labeled cells demonstrating the colocalization of γ-tubulin and PYK-2 at the uropod of NK cells.