FAK-WT-GFP, FAK-Y397F-GFP and FAK-I936E I998E-GFP were obtained as previously described [17 (link)]. The LD2-LD4-GFP sequence of paxillin (aa 136–296) was inserted into the pEGFP–C1 plasmid (Clontech, Kusatsu City, Japan), creating a fusion of enhanced GFP (EGFP) to the N terminus of paxillin. Melanoma cells were co-transfected with siRNA and 4 µg FAK WT or FAK mutant DNA or with LD2-LD4-GFP DNA 4 µg using lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), according to the manufacturer’s instructions. Transfected cells were incubated for 24 h at 37 °C before use.
Free full text: Click here
