Fig. 4.
Reducing EGFR-induced cyclin D1 promoter activity by interruption of EGFR–RHA Interaction. (A) Schematic of RHA deletion constructs. +, binding to EGFR; −, no binding to EGFR. RGG, RHA RGG domain. (B) Disturbance of EGFR–RHA interaction by deleting the helicase domain of RHA. HEK293T cells were cotransfected with indicated plasmids followed by cell lysis, immunoprecipitation, and Western blot. Expression levels of whole-cell lysates blotted for RHA (Flag), EGFR, and tubulin are shown. IB, immunoblotting; IP, immunoprecipitation; WCL, whole-cell lysate. (C) Reduction of EGFR-induced cyclin D1 promoter activity by deleting the helicase domain of RHA. Luciferase assay was performed in HeLa cells with stable knockdown of endogenous RHA. (D) Reduction of EGFR–RHA interaction by EGFRmNLS. HEK293T cells were transfected with indicated plasmids and lysed for immunoprecipitation-Western blot. (E) Abrogation of EGFR/RHA-induced cyclin D1 promoter activity by EGFRmNLS.