Fig. 4

RICTOR binds FLNA. a FLAG-RICTOR is pulled down after lysing U87vIII cells with CHAPS or Triton X-100 as a detergent in lysis buffer. An immunoblot shows that RICTOR is associated with mTOR and FLNA in samples from CHAPS-treated extracts but only with FLNA in Triton X-100-treated extracts b Immunofluoresence labeling of two individual U87vIII cells grown in normal condition. Majority of FLNA and RICTOR colocalize along cell membrane and cytoplasm. Anti-FLNA antibody and Anti-RICTOR antibody were used to stain FLNA and RICTOR, respectively (Exposure time: FLNA 334 ms; RICTOR 540 ms; DAPI 41 ms). However, only RICTOR is found in nuclei of the cells. Overall results indicate that FLNA is an mTORC2 associated protein which is a binding partner of RICTOR