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. 2015 Nov 3;6(39):42183–42196. doi: 10.18632/oncotarget.5619

Figure 1. MEK inhibition blocks cell growth in HRAS mutant cells.

Figure 1

A. Presentation of HRAS mutant and HRAS wild-type cell lines. Mutations were detected by PCR of cDNA covering the open reading frame of HRAS and confirmed mutation status from COSMIC data base. PCR products were purified and sent for sequencing. B. Distribution of HRAS mutations from tumor tissue of lung (upper panel) and bladder (lower panel) available from COSMIC data base. C, D. Mutant HRAS und wild-type HRAS cell lines were treated with 6 increasing concentrations of AZD6244 and MEK162 for 96 hours. Then, cell growth was measured by Cell Titer Glo according to the manufacturer's instructions. EC50 values were calculated with GraphPad Prism and depicted at bars. Statistical significance between mutant and wild-type cell lines was calculated with student's t-test. E. All cell lines were kept under equal conditions, then treated with 500nM of AZD6244, MEK162 and PD0325901 for 1 hour and next lysed and subjected to Western blot. Phosphorylation levels of ERK and MEK were detected by specific anti-phospho antibodies. Loading was verified by specific antibodies to total ERK, MEK and anti–actin.