Fig. 3.

Co-immunoprecipitaion of Rabin8 and Rheb overexpressed in HEK293 cells after transfection with proper vectors. The levels of Rabin8, tuberin, mTOR and β-аctin were measured using immunoblot analysis with specific antibodies. (a) The expression levels of Rabin8, Myc-Rheb, and mTOR after co-transfection with control vectors pcDNA3.1 and pCMVTag3A ( lane 1 ) or Rabin8 and Myc-Rheb ( lanes 2–4 ) in the presence of growth factors ( lanes 1,2 ), absence of growth factors ( lane 3 ), and after 15-min stimulation of cells grown in media without growth factors with media containing growth factors ( lane 4 ). (b) Co-immunoprecipitation using the lysates from Fig. 3a with antibodies against Rabin 8 ( lanes 1–4 ) or control rabbit IgG ( lane 5 ) and immunoblot analysis with antibody against Myc. The lysate from Fig. 3a ( lane 2 ) was used for the co-immunoprecipitation with control rabbit IgG antibody. (c) Co-immunoprecipitation using the lysates from Fig. 3a with antibodies against Myc and immunoblot analysis with antibody against Rabin8, mTOR, and Myc.