FIG. 3.
Interaction of Ha-Ras with TPR1 and facilitation of the interaction by Gα16. (A) HEK293T cells were transiently transfected with the FLAG-TPR1 construct either alone or in combination with expression vectors for Ras and/or Gα16 as indicated. The cell homogenates were subjected to immunoprecipitation with an anti-FLAG agarose affinity gel and analyzed by Western blotting. Total FLAG-TPR1 in the immunoprecipitates (IP) was shown by probing the Western blot (WB) with an anti-FLAG MAb (middle panel). The IP (top two panels) and cell homogenates (H) (bottom two panels) were probed with anti-Ras and anti-Gα16 sera to assay for the abilities of Ras and Gα16 to coimmunoprecipitate with FLAG-TPR1 and for their expression in the cells. Results are representative of at least three independent experiments. (B) FLAG-TPR1 was transiently expressed in HEK293T cells and used for coimmunoprecipitation (IP) of Ras with or without exogenous Raswt. Samples were analyzed by Western blotting.