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. 2013 Nov 22;8(11):e79763. doi: 10.1371/journal.pone.0079763

Figure 3. Analysis of the focal adhesion number and FAK levels in CD and controls fibroblasts before and after P31-43 treatment.

Figure 3

A. Increase of focal adhesion number in CD, as revealed through FAK staining before and after P31-43 treatment. a) Confocal immunofluorescence images of fibroblasts from CD patients and controls stained with antibodies against FAK. Representative fields. b) Confocal immunofluorescence images of fibroblasts stained with antibodies against FAK from CD patients and controls treated with P31-43 for 30 min. Representative fields. c) Statistical analysis of the number of FAK-positive focal adhesions per cell before and after P31-43 treatment. Focal adhesions of 30 fibroblasts of several fields from 3 independent experiments with 6 patients and 6 controls were counted. Columns represent the means, and the bars represent the standard deviations. Student's t-test. * = p<0.05; ** = p<0.01; *** = p<0.001. B. FAK protein levels are increased in fibroblasts from CD patients with respect to the controls. a) Western blot analysis of fibroblast protein lysates from controls and CD patients. The upper line is stained for FAK, and the lower line is stained for tubulin. Representative experiment. b) Densitometric analysis of western blots stained for FAK in fibroblasts from 5 CD patients and 5 controls. FAK levels were normalized to the tubulin levels. Columns represent the means, and the bars represent the standard deviations. Student's t-test. *** = p<0.001. C. FAK phopshorylation is increased in CD fibroblasts and is further increased after P31-43 treatment for 30′. a) Western blot analysis of fibroblasts proteins lysates from controls and CD patients treated or not with P31-43. The upper line is stained for pY-FAK, the middle line is stained for FAK and the lower line is stained for tubulin. Representative experiment. b) Densitometric analysis of western blots stained for pY-FAK and FAK in fibroblasts from 5 CD patients and 5 controls. pY-FAK levels were normalized to the FAK levels. Columns represent the means, and the bars represent the standard deviation. Student's t-test. * = p<0.05; *** = p<0.001.