Fig. 1

RHEB interacts with BRAF, decreasing BRAF/CRAF heterodimerization and RAF/MEK/ERK signaling. a Immunoprecipitation of FLAG-RHEB from HEK 293T cell lysate, and Western blot of CRAF, BRAF, and FLAG. b FLAG tagged RHEB WT, T38A, and D60I were expressed in HEK 293T cells. Cell lysates were collected and anti-FLAG immunoprecipitation was performed followed by Western blot for BRAF and FLAG. c Western blot showing levels of RHEB, phosphorylated ERK, total ERK, phosphorylated S6K, total S6 K and ACTIN in normal and RHEB knockdown HEK 293T cell lines. Compare the amount of phosphorylated ERK relative to the total amount of ERK. d Western blot showing levels of RHEB, phosphorylated ERK, total ERK and ACTIN in normal and overexpressed-RHEB HEK 293T cell lines. Compare the amount of phosphorylated ERK relative to the total amount of ERK. e Endogenous BRAF was immunoprecipitated from HEK 293T cells containing endogenous RHEB or overexpressed RHEB. Western blot of RHEB, CRAF, and BRAF is shown