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. 2014 Feb;34(3):348–361. doi: 10.1128/MCB.00825-13

FIG 7.

FIG 7

Differential abilities of Grb2 domain mutants to rescue paxillin expression, FAK autophosphorylation at Tyr397, and PTPα-Tyr789 phosphorylation. (A) Schematic diagram of siRNA-resistant myc-tagged Grb2-WT/mutants. Silent mutations were made in the depicted region targeted by the Grb2 siRNA. The closed circles depict the domain with the inactivating point mutation. (B to E) MEFs treated with either control (Ctl) or Grb2 siRNA duplex or cotransfected with Grb2 siRNA and Grb2-myc-WT or mutants were kept in suspension for 1 h and replated on FN-coated dishes or coverslips for 15 min. (B) Paxillin expression was analyzed by probing cell lysates, and the graph in panel C shows the relative amount of paxillin per unit of FAK as calculated from densitometric analysis of 3 experiments as in panel B. (D) FAK-Tyr397 and PTPα-Tyr789 phosphorylation was determined by probing cell lysates (D), and the graph in panel E shows the relative amount of FAK-phospho-Tyr397 per unit of FAK as calculated from densitometric analysis of 3 experiments as in panel D.