Fig. 6.

Mechanistic role of MRAS in H/N/KRAS-less paradoxical ERK activation. (A) H/N/KRAS-less MEFs with or without Mras were transiently transfected with the indicated Flag-tagged Mras constructs. Twenty-four hours later, cells were treated with 0, 0.01, 0.1, or 1.0 µM LY3009120 for an additional 24 h. Lysates were collected and immunoblotted with indicated antibodies. Representative blots from three experiments that show similar results are shown. (B) MEFs were treated with LY3009120 for 24 h, and lysates were collected for coimmunoprecipitation with the MRAS antibody, followed by immunoblotting with the BRAF and CRAF antibody. Input lysates were immunoblotted with indicated antibodies. Representative blots from two experiments that show similar results are shown.