Fig. 5
From: ERK5 is activated by oncogenic BRAF and promotes melanoma growth
Oncogenic BRAF increases ERK5 nuclear localization and transcriptional transactivator activity. a Nucleo-cytoplasmic fractionation in HEK-293T cells transfected with equimolar amounts of wt ERK5 in combination with the empty vector pCAG, constitutively active MEK5 (MEK5DD) or BRAFV600E plasmids. BRAFV600E increases the level and the phosphorylation of nuclear ERK5. GAPDH and Lamin B1 were used as cytoplasmic or nuclear markers, respectively. b Nucleoplasm and chromatin-bound fraction from HEK-293T cells transfected with equimolar amounts of empty or BRAFV600E pcDNA plasmids in presence or not of wt ERK5. Rb and Histone H4 were used as nucleoplasm or chromatin-bound markers, respectively. (a, b) Representative blots from three independent experiments. c Quantification of dual reporter luciferase assay in M26c melanoma cells showing that BRAFV600E enhances the transcriptional transactivator activity of wt ERK5. Relative luciferase activity was firefly/Renilla ratios, with the level induced by control equated to 1. Data represent mean ± SD of three independent experiments. **p < 0.01, ***p < 0.001 as determined using one-way ANOVA
