Fig. 2.

Gas plasma treatment modulated signaling and scaffolding proteins that link the integrin receptors to the actin cytoskeleton. (a) Gene expression levels of focal adhesion kinase (FAK), paxillin α (PXNα), vinculin (VCL), and talin (TLN1) were determined by qPCR. Scheme of focal adhesion complexes with associated protein targets. (b, d) Protein quantification of the non-phosphorylated proteins and activated pFak, pPxn, pVcl as well as tyrosine kinase (Src) and phosphoinositide-3-kinase (PI3K) were determined by Western blot analysis in relation to incubation time with gas plasma-treated medium as indicated. Data were normalized to GAPDH and untreated controls (ctrl) at indicated time points (n > 5) and presented as mean +SE. Statistical analysis was done by unpaired two-tailed Student's t-test with significances of *p < 0.05, **p < 0.01, and ***p < 0.001. (c, e) pDFs were grown on glass coverslips, incubated with gas plasma-treated medium (pl-20 s), fixed, and subjected to fluorescent staining for determination of expression and distribution of FA proteins (red) such as anti-Fak (I), anti-Pxn (II), anti-Vcl (III, arrowheads show vinculin positive cellular protrusions), and their phosphorylated counterparts in (e) with anti-pY397-FAK (I), anti-p-Pxn (II), and anti-pVcl (III). The cell nuclei were stained with DAPI (blue). Scale bars are 50 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)