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. 2006 Sep 11;174(6):863–875. doi: 10.1083/jcb.200603013

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Copyright © 2006, The Rockefeller University Press

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Figure 2. — Expression of shRNA depletes fascin in B16F1 cells. (A) Target sequences for RNAi. Tc (left) is common for mouse and human fascin; Th and Tm (right) are common for human and mouse fascin mRNA, respectively, and differ in two base pairs. (B) Immunoblot analysis. Lysates prepared from FACS-purified cell populations 5 d after transfection were probed with fascin antibody. Tubulin served as a loading control. (C) Microscopic analysis. GFP fluorescence serves as a marker for cells transfected with the Tm, Th, or Tc constructs. Fascin is visualized by immunofluorescence and actin is stained with fluorescent phalloidin. Merged images show GFP (green) and fascin staining (red) simultaneously. Intensity line scans (right) through one transfected and one untransfected cell (lines in merged images) show fascin and GFP intensities in red and green, respectively. Bar, 20 μm.