Fig. 4

α2 integrins suppress Src and Shp2 activity to activate RhoGDI at junctions. a Western blot of WT monolayers in Ca²⁺ treated with either DMSO or BTT (20 μm, 1h) probed for pY416-Src and total Src and quantification from 4 independent experiments. b Images of WT monolayers in Ca²⁺, treated with either DMSO or BTT (20 μm, 1h), fixed and stained for p-Src and E-cadherin and quantification of junctional p-Src. Data are from at least 30 images per condition and over 3 independent experiments. c Images of WT monolayers in Ca²⁺ , treated with either DMSO or BTT (20 μm, 1h) and/or PP2 (1 μm, 1h), fixed and stained for E-cadherin and quantification of junctional E-cadherin. Data are from at least 30 images per condition and over 3 independent experiments. d FRET/donor ratiometric images of live control and α2KD cells expressing the Cdc42 GDI FRET biosensor with or without BTT (20 μm, 1h) or PP2 (1 μm, 1h) treatment. e Quantification of FRET ratio levels from cells as in d. Data are from at least 18 images per condition and over 3 independent experiments. F Western blot of WT monolayers in Ca²⁺ treated with either DMSO or BTT (20 μm, 1h) probed for pY542-Shp2 and total Shp2 and quantification from 4 independent experiments. g Images of WT monolayers in Ca²⁺, treated with either DMSO or BTT (20 μm, 1h), fixed and stained for pY542-Shp2 and E-cadherin. h Quantification of junctional pY542-Shp2 as in g. Data are from at least 35 images per condition and over 3 independent experiments. i Images of WT monolayers in Ca²⁺ , treated with either DMSO or BTT (20 μm, 1h) and/or shp099 (1 μm, 1h), fixed and stained for E-cadherin. j Quantification of junctional E-cadherin as in i. Data are from at least 30 images per condition and over 3 independent experiments. Scale bars 10μm throughout.; n.s. not significant, ***p<0.001, **p<0.01, *p<0.05