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. 2004 Sep 27;166(7):1081–1091. doi: 10.1083/jcb.200402138

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Copyright © 2004, The Rockefeller University Press

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Figure 4. — Resting CD44 ⁺ COS-7 transfectants show Smad1 cytoplasmic sequestration; BMP-7 treatment initiates Smad1 nuclear translocation. Parental COS-7 cells, which lack CD44 expression, exhibit diffuse staining of Smad1 protein (red fluorescence) throughout the cytoplasm and the nucleus (A). No change in the diffuse staining pattern of Smad1 protein was observed in parental COS-7 cells after treatment with BMP-7 (B). Stable COS-7 transfectants, expressing cell surface CD44, exhibited Smad1 immunolocalization preferentially to the cytoplasm, and Smad1 staining was nearly absent from the nucleus (C). After treatment of the CD44⁺ COS-7 transfectants with BMP-7, the nuclear translocation of Smad1 protein was observed (D). Insets in all panels show two-color fluorescence with the DAPI nuclear stain of the same cells at lower magnification. Bar, 20 μm for the main panels.