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. 2012 Apr 10;7(4):e33085. doi: 10.1371/journal.pone.0033085

Figure 5. Inhibition of Src family kinases decreases cell migration and phosphorylation of caveolin-1 on tyrosine-14. (A).

Figure 5

MDA-MB-231 cells stably transduced with shRNA either targeting Luciferase (sh-ctrl) or caveolin-1 (sh-cav1) were pre-incubated with DMSO (D) or 10 µM PP2 for 30 minutes, wounded with a pipet tip, stimulated (+) or not (-) with 3% of serum and images were recorded at 0 and 16 hours post-wounding. The wounded area was measured with the Image J software and the percentage of wound closure was plotted (upper panel). Total protein extracts were prepared and analyzed by Western blotting with antibodies against caveolin-1, pY14-caveolin-1 and actin. (B) Confluent monolayers of B16-F10 cells transfected with either pLacIOP or pLacIOP-caveolin-1 were pre-incubated with 1 mM IPTG for 24 hours. Then, cells were treated with DMSO (D) or PP2, wounded with a pipet tip, stimulated with 3% of serum and analyzed at 0 and 7 hours post-wounding, as described in (A). Total protein extracts were prepared and analyzed by Western blotting. Values in (A) and (B) were averaged from 3 independent experiments (mean ± SEM). *P<0.05.