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. 2014 Oct 1;127(19):4308–4321. doi: 10.1242/jcs.155879

Fig. 8.

Fig. 8.

Kindlin-2-ILK interaction is required for β1 integrin activation. (A) Immunoblot (IB) analysis of kindlin-2 and vinculin in parental, control (scrambled, SC) and kindlin-2-knockdown (K2 shRNA) NIH3T3 cells. (B) Immunofluorescence staining of endogenous kindlin-2 and vinculin in scrambled control and kindlin-2-knockdown NIH-3T3 fibroblasts. Spread kindlin-2-knockdown cells were selected for imaging. (C) Subcellular localization of GFP or GFP-tagged kindlins, and of endogenous vinculin in K2 shRNA cells plated on fibronectin for 5 h. (D) β1 integrin surface levels measured by flow cytometry. (E) Integrin α5β1 activation index, measured by flow cytometry and normalized to β1 staining (n = 4). (F–H) Scramble control or kindlin-2-knockdown fibroblasts were transfected with GFP or GFP-tagged kindlins and α5β1 integrin activation and β1 surface levels was assessed in transfected cells. Bar graphs represent mean±s.e.m.; n≥3; *P<0.05; **P<0.01; ***P<0.001; ns, not significant (Student's t-test). Scale bars: 10 µm.