Sin1 isoforms except Sin1δ form mTORC2 complex.

<p>A. Rictor pulled down Sin1α, Sin1β, Sin1γ, but not Sin1δ. HEK-293T cells were transiently transfected for 24h with HA-Sin1 isoform plasmids. Cell lysates (left-hand side) and rictor immunoprecipitates (right-hand side) were analyzed for mTOR, rictor and HA by western blotting. B. Sin1α, Sin1β, Sin1γ, but not Sin1δ could pull down rictor and mTOR. HEK-293T cells were transiently transfected for 24h with HA-Sin1 isoform plasmid respectively. Cell lysates (left-hand side) and HA immunoprecipitates (right-hand side) were analyzed for mTOR, rictor and HA by western blotting. All experiments were repeated for three times with the same results.