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Mitochondrial ERK localization depends on differential phosphorylation.

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posted on 2018-02-28, 18:40 authored by Katia E. Helfenberger, Nerina M. Villalba, Bruno Buchholz, Alberto Boveris, Juan José Poderoso, Ricardo J. Gelpi, Cecilia Poderoso

Representative confocal images of LP07 tumor lung transfected cells with wild type ERK/V5 WT (left panel), Y183A/V5 (middle panel) and T183A/V5 (right panel). Forty-eight hours post transfection, cells were incubated with 1 μM H2O2 during 0–30 min. From top to bottom of the panels images show increasing H2O2 incubating times. Mitochondria were visualized in red by staining with Mitotracker Deep Red. Cells were fixed and incubated with anti-V5 antibody and a secondary antibody conjugated with Cy2, and analyzed in an Olympus FV1000 confocal microscope. Co-localization was highlighted in green on the grayscale images (last column of each panel). Images directly exported from Olympus Fluoview acquisition program. Fold changes relative to control images of combined MitoTracker and Cy2 intensity for ERK2 WT (Panel A), Y185A (Panel B) and T183A (Panel C). Basal conditions without H2O2 arbitrarily defined as 1. Bar = 10 μm.