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. Author manuscript; available in PMC: 2015 Mar 1.
Published in final edited form as: Sci Signal. 2014 Jun 17;7(330):ra57. doi: 10.1126/scisignal.2004838

Figure 1. FAK/Src-mediated Cas phosphorylation transduces ECM stiffness into mechanosensitive cell cycling.

Figure 1

Serum-starved MEFs were incubated on low or high stiffness hydrogels and stimulated with FBS as indicated. Total cell lysates were immunoblotted. (A–C) Lysates analyzed for phosphorylated and total Cas (A), paxillin (B; Pax), or vinculin (C; Vinc). N=3 independent biological replicates for (A) to (C). (D) Starved MEFs transfected with a control (ctrl) siRNA or siRNAs to Cas, paxillin, or vinculin were seeded on high stiffness hydrogels with FBS for 20 hours. S phase entry was assessed by EdU incorporation. Error bars show mean ± SD, N=4 independent biological replicates. (E) Same experiment as in (A) but lysates were probed for Tyr397, Tyr576, Tyr861, and total FAK. (F) Starved MEFs infected with adenoviruses (FRNK or FAK397F) were replated on stiff hydrogels with FBS for 9 hours and probed for phosphorylated and total Cas. The vertical white bar indicates removal of irrelevant lanes from a single blot. N=3 independent biological replicates. (G) WT MEFs, SYF-null (MEFs deficient in Src, Yes, and Fyn) and c-Src-reconstituted SYF-MEFs were plated on high stiffness hydrogels and stimulated with FBS for 9 hours. N=3 independent biological replicates for (E) to (G).