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

FIG 1.

FIG 1

Grb2 knockdown reduces PTPα Tyr789 phosphorylation. (A) Left panel, wild-type (PTPα+/+) MEFs were starved overnight (adherent cells [Adh]) and then trypsinized and kept in suspension for 1 h (0) before plating on FN-coated plates for 30 min (30). Right panel, PTPα-null MEFs (PTPα−/−) and PTPα-null MEFs reexpressing WT or Y789F-PTPα were stimulated on FN as described above. PTPα immunoprecipitates were probed for Grb2, phosphotyrosine (PY20), and PTPα. (B) Wild-type MEFs were treated with control (Ctl) or Grb2 siRNA duplex 1 or 2 for 24, 48, and 72 h. Cell lysates were probed for Grb2, actin, PTPα-pY789, and PTPα. (C) Wild-type MEFs were treated with control (Ctl) or Grb2 siRNA for 48 h and then starved overnight. Cells were trypsinized and held in suspension for 1 h (0), and then a portion of the suspended cells were plated on FN-coated plates for 15 min (15). Lysates were probed as shown. (D) Wild-type MEFs treated with either control (Ctl) or Grb2 siRNA or cotransfected with Grb2 siRNA and myc-tagged Grb2-WT were stimulated on FN-coated dishes for 15 min. Lysates were probed as shown.