Fig. 2. p85β regulates cell adhesion

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Fig. 2. p85β regulates cell adhesion Fig. 2. p85β regulates cell adhesion.(A) p85α or p85β expression was analyzed in WB in stably transfected NIH3T3 cells. p85β regulates cell adhesion.(A) p85α or p85β expression was analyzed in WB in stably transfected NIH3T3 cells. Freshly isolated MEF (from WT, p85α−/− or p85β−/− mice) were genotyped by PCR. A fraction of the cells were activated with PDGF (50 ng/ml) and used in an adhesion assay, alone or on fibronectin (FN), or in a migration assay on transwells coated with a thin layer of FN or collagen (COL)(10 µg/ml). Graphs show the percent of response relative to the maximal in control cells (100%; mean ± s.d.; n  =  5). (B) Control and p85β-overexpressing NIH3T3 cells were transfected with FAK siRNA (48 h) or preincubated with FAK inhibitor (10 µM PF-573228) for 1 h at 37°C prior to stimulation with 50 or 100 ng/ml PDGF (indicated) on FN. The cells were assayed in an adhesion assay (as in A). Some of the cells transfected with siRNA or treated with the inhibitor were lysed and lysates examined in WB using were anti-FAK Ab or anti-pTyr576/577-FAK Ab (indicated). n.s. =  non-statistical significance; ** P<0.01, * P<0.05; Student's t-test. Ariel E. Cariaga-Martínez et al. Biology Open 2014;bio.20148185 © 2014. Published by The Company of Biologists Ltd