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. 2007 Sep 10;178(6):981–994. doi: 10.1083/jcb.200703106

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Copyright © 2007, The Rockefeller University Press

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Figure 4. — Imp7 and 8 are required for nuclear accumulation of BMP-activated Smad1 in HeLa cells. (A) HeLa cells transfected with indicated siRNA duplexes (40 nM) were analyzed for mRNA levels of Imp7 or Imp8 by real-time RT-PCR. GAPDH was used as the internal standard for quantitation. Error bars indicate SD. (B) HeLa cells transfected with siRNAs were treated with BMP2 (100 ng/ml, 1 h) as indicated and analyzed by immunoblotting using antibodies specific for phospho-Smad1 or total Smad1. (C) HeLa cells transfected and treated as in B were immunostained with the PS1 antibody recognizing phospho-Smad1 and analyzed by fluorescence microscopy. Bar, 10 μm. (D) BMP2-induced transcriptional up-regulation of Smad6 was inhibited by siRNA targeting Imp7 or Imp8. HeLa cells transfected and treated as in B were analyzed for Smad6 mRNA level by real-time RT-PCR, with GAPDH serving as the standard. The plotted data are derived from three experiments and the error bars indicate SD.