Figure 7
From: Activation state-dependent interaction between Gαq subunits and the Fhit tumor suppressor
Fhit does not affect G q -mediated TPR1 interaction or Ras activation. A, HEK293 cells were co-transfected with different combinations of pcDNA3 (Vector), Fhit, Gα16, Gα16QL, Flag-TPR1 and Flag tag constructs. One day later, cells were immunoprecipitated with anti-Flag agarose affinity gel and probed for the presence of Fhit, Gα16, and Flag-TPR1 in the immunoprecipitates using specific antibodies as indicated. B, HEK293 cells were co-transfected with either pcDNA3 (Vector) or Fhit in combination with pcDNA3, Gα16 or Gα16QL, and subjected to Ras activation assay. The results shown on the bar chart represent relative band intensities of Ras from RBD-Raf-1 immunoprecipitation normalized against their corresponding amounts of total Ras (RBD-Raf-1/TCL). The value of vector-transfected basal control was set as 100%. * Transfection of Gα16QL significantly induced Ras activity as compared to vector control (Dunnett’s t test, P < 0.05). Immunoblots shown represent one of three sets; two other sets yielded similar results.