LY3009120 induces ERK target gene transcription and cell proliferation in H/N/KRAS-less MEFs. (A) Total RNA was extracted from parental and H/N/KRAS-less MEFs treated with 0, 0.003, 0.03, 0.3, 1.0, or 3.0 µM LY3008120 for 24 h. Levels of Ccnd1, Dusp4, Dusp6, Spred2, Spry2, Etv1, Etv4, and Etv5 transcript were examined using quantitative RT-PCR. Results are shown as fold-change relative to DMSO-treated control represented by gray shading at a fold-change of 1.0. Error bars represent SEs from a representative experiment of three experiments. (B) Parental and H/N/KRAS-less MEFs were seeded and treated 24 h later with 0.0, 0.001, 0.01, 0.1, or 1.0 µM LY3009120 in serum-free media. Cell numbers were monitored for an additional 4 d using the IncuCyte Live-Cell Analysis System. Error bars represent SDs from a representative experiment of two experiments.