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. 2004 Dec;15(12):5635–5646. doi: 10.1091/mbc.E04-06-0490

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Copyright © 2004, The American Society for Cell Biology

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Figure 8. — Ccn2-/- mouse embryonic fibroblasts show impaired spreading on fibronectin. Ccn2+/+ and Ccn2-/- mouse embryonic fibroblasts (MEFs) were allowed to adhere to fibronectin for 40 min as described in Materials and Methods. In addition, Ccn2-/- fibroblasts were preincubated with rCCN2 for 1 h before adhesion to fibronectin. Cells were fixed in paraformaldehyde and subjected to indirect immunofluorescence analysis with anti-α-SMA antibody and detected with an appropriate FITC-conjugated antibody as described in Materials and Methods and in the legend to Figure 1C. Forty minutes postadhesion, Ccn2+/+ MEFs adhered to fibronectin show extension of filopodia and α-SMA localization to the cell periphery (arrow with f, Ccn2+/+) and display a few stress fibers (arrow with a, Ccn2+/+). Conversely, Ccn2-/- MEFs do not display filopodia, α-SMA localization to the cell periphery, or stress fibers (Ccn2-/-). Ccn2-/- fibroblasts that were preincubated with rCCN2 for 1 h before adhesion to fibronectin showed extension of filopodia and α-SMA localization to the cell periphery (arrow with s, Ccn2-/-) and display a few stress fibers (arrow with a, Ccn2-/- + rCCN2). Nuclei were detected by DAPI staining (blue).