Figure 4.

A–C) HGFs were transfected with control (scrambled) siRNA or talin-1 and talin-2 siRNAs for 48 h, as outlined in Materials and Methods. Cells were trypsinized and replated on FN for 3 h in medium containing 2% serum. After treatment with vehicle or with IL-1β (20 ng/ml) in serum-free medium for 5, 15, or 30 min, cell lysates were immunoblotted for talin-1, talin-2, and GAPDH (A); for the indicated proteins in focal adhesion preparations (B); and for phospho-ERK and total ERK (C, left panel). Histogram (C, right panel) shows densitometry data from immunoblots of C. D) NIH 3T3 cells were transfected with control (scrambled) or kindlin-2 siRNA using DharmaFect Reagent 1 for 48 h, as outlined in Materials and Methods. Cells were replated on FN for 3 h in medium containing 2% serum. After treatment of cells with vehicle or IL-1β (20 ng/ml; 5, 15, or 30 min) in serum-free medium, cell lysates were immunoblotted for kindlin-2 and GAPDH. E) Focal adhesion proteins from the experiments in D were prepared and immunoblotted for the indicated proteins. F) In the kindling-2-knockdown experiments, phospho-ERK and ERK were immunoblotted (left panel), and the densitometry results were quantified (right panel).