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Fig. 3 | Cell Communication and Signaling

Fig. 3

From: Syndecan-4/PAR-3 signaling regulates focal adhesion dynamics in mesenchymal cells

Fig. 3

PAR-3 silencing impaired Thy-1-accelerated microtubule-dependent focal adhesion disassembly. a DI TNC1 cells treated with Nocodazole and washout were stimulated or not (NS) with Thy-1-Fc-Protein A (Thy-1) or TRAIL-R2-Fc-Protein A (TRAIL) for different time periods. FAs were detected by immunofluorescence staining for vinculin (red) and nuclei (blue). Scale bar = 10 μm. Graphs show quantification of the FA number per cell (left panel) and average area of FAs (right panel) observed in DI TNC1 astrocytes after the indicated treatments. Values in the graphs represent the mean ± s.e.m. determined from at least 50 cells in each experimental condition per experiment. b DI TNC1 astrocytes co-transfected with mCherry-vinculin and siRNA (control or PAR-3) were treated with Nocodazole and washout. Cells were then stimulated as in (a), and a 30-min time-lapse microscopy video was recorded. Values in the graphs represent the mean ± s.e.m. determined from scoring at least 10 mCherry-positive focal adhesions per experiment. c Model depicting the interaction of αvβ3 integrin and Syndecan-4 with Thy-1, which triggers signaling events that include activation of FAK and PI3K generating PIP3. An unidentified Rac1 GEF, is then recruited to the plasma membrane, where it activates Rac1 and leads to cell migration. d DI TNC1 astrocytes, transfected as in (a), were treated with Nocodazole and washout, and cells were stimulated for different time periods (0–15 min). Cell lysates were immunoblotted for p-FAK or total FAK. e Graph values are compared against the normalized time 0 and represent the average numbers obtained from the ratio of p-FAK/FAK densitometric data analysis. All graphs show the mean ± s.e.m. of n = 3. Significant differences compared to control conditions are indicated as *P < 0.05, **P < 0.01, and ***P < 0.001

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