Fig. 4

Expression of LD2-LD3-LD4 blocks kinase-independent, scaffolding functions of FAK at FAs a-b) Super resolution images and quantification of the FA-localization of GFP Src_dSH2 in control (GFP Src_dSH2 only) and mCherry LD2-LD3-LD4 expressing cells (marked with asterisk). Cells were fixed with PFA and immunostained for Vinculin. Expression of mCherry LD2-LD3-LD4 leads to 84.3% decrease in the Src_dSH2/Vinculin ratio compared to control cells (100 ± 6.68, n = 419 FAs from 24 control, 15.67 ± 0.93, n = 373 FAs in 22 mCherry LD2-LD3-LD4 expressing cells). c-d) Confocal images (c) and quantification (d) of control (top panel), LD2-LD3-LD4 expressing (intermediate panel) and PF228 treated (bottom panel) cells, fixed with methanol/acetone and immunostained for p130Cas and Talin. Expression of LD2-LD3-LD4 leads to a 68% decrease in the p130Cas/Talin ratio compared to control cells, whereas treatment with PF228 does not elicit any significant change (100 ± 3.56, n = 505 FAs from 33 control, 31.89 ± 2.46, n = 564 FAs from 35 LD2-LD3-LD4 expressing and 100.2 ± 5.6, n = 515 FAs from 30 PF228-treated cells). F Scale bars: 10 μm. The error bars represent standard error of the mean (S.E.M). ***; p < 0.0001