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. Author manuscript; available in PMC: 2015 May 1.
Published in final edited form as: Mol Cancer Res. 2014 Feb 11;12(5):795–802. doi: 10.1158/1541-7786.MCR-13-0581

Fig. 3. Phospho-ERK1/2 levels are hyperactivated in PRT #3 and PRT #4 cell lines after RAF inhibitor withdrawal.

Fig. 3

(A) Parental 1205Lu cells were seeded overnight in drug-free medium, and PRT #3 and PRT #4 cells were seeded in 1µM PLX4720. The next day, cells were washed and then administered either 1µM PLX4720 or DMSO for a 16 hour treatment. Western blots were performed to assay ERK1/2 activation status. (B) PRT #3 and PRT #4 cells were cultured in PLX4720 (1µM) and then medium changed to drug-free medium. Cell lysates were harvested at the indicated time and analyzed by western blotting for levels of phosphorylated MEK as well as phosphorylated ERK1/2. (C) Similar to (B), cells were lysed after culturing in drug-free medium for the indicated time, and the CDK inhibitors p21Cip1 and p27Kip1 were analyzed by western blot, as well as the putative ERK target sprouty2.