Figure 3. Src-focal adhesion kinase (FAK) signalling regulates cell migration and invasion.
A. Relative mRNA levels of PTK2/FAK and Src and, B. protein levels of total FAK, Src, paxillin, pSrc-Y416, pFAK-Y397, pPaxillin-Y118 and β-actin after transient knockdown by Scram (control), FAK and Src siRNA molecules in late passage human foreskin keratinocytes (HFK) expressing HPV16 E6/E7 genes. C. Representative phase contrast images of scratch wound assay on collagen-I coated plates showing cell migration after transient knockdown with scram, FAK and Src siRNA in the late passage E6/E7-HFK, which was D. quantified and represented as μm2/hr. E. H&E staining and immunofluorescence detection of BrdU uptake and nuclei (DAPI) on 3D-organotypic rafts established from late passage E6/E7-HFK seeded on retinoblastoma-depleted HFF embedded collagen-I plugs and treated with specific inhibitors of FAK (PF573228) and Src (dasatinib) activities. Rafts treated with DMSO served as control. Invasive incidents are indicated by arrows. F. Quantification of number of invasive incidents across the rafts per cm. Scale bars represents 100 μm. N = 3 independent experiments, mean ± SEM, **p < 0.01 compared to the respective controls, *p < 0.05 compared to the control.