ERK2 is compartmentalized upon EGF stimuli in LP07 tumor lung cells.

<p>LP07 cells were transiently transfected with wild type ERK2-V5 (wt), using Lipofectamine 2000 reagent. Twenty-four hours before the experiment, serum was removed from the culture media to avoid any impact on ERK phosphorylation and localization due to serum effect. Forty-eight hours post-transfection, LP07 were stimulated with 10ng/ml EGF peptide for the indicated times. Temporal activation and distribution of ERK2-V5 in the subcellular fractions were analyzed by western blot using antibodies against V5 tag. Representative images of three independent experiments (Panel A). Protein loading was determined with antibodies anti-TOM40 for mitochondria, actin for cytosol, and polymerase II (POLII) for nuclei. In Panel B, the graph indicates amounts of ERK2-V5 respective to control cells (100%). Panel C shows a representative immunoblot with specific antibodies for each subcellular fraction (M = mitochondria; C = cytosol; N = nucleus).