Figure 4: Pdlim5 phosphorylation was blocked in AMPKα1/2-null MEFs.
From: Augmented AMPK activity inhibits cell migration by phosphorylating the novel substrate Pdlim5
(a) WT or AMPKα1/2-null MEFs were stimulated with AICAR (1 mM) for 15 min. TCLs were subjected to immunoblotting with indicated antibodies. (b) Phase-contrast images of WT-MEFs and AMPKα1/2-null MEFs were collected during the scratch assay in the presence or absence of AICAR (1 mM). Each panel shows analysis of migration paths over 8 h. The origins of migration of each cell were superimposed at [0, 0]. (c) Bar graph showing the migrating speed of each cell (from b). (d) Bar graph showing migration directionality of each cell (from b). Numbers in the bars indicate n. Data are representative of means±s.e.m. from three independent experiments. Significance of differences between series of results was assessed using one-way analysis of variance, followed by a post-hoc comparison with Dunnett’s method for multiple comparisons. *P<0.01 compared with WT without AICAR treatment. (e) Immunostained images of WT-MEFs and AMPKα1/2-null MEFs in the presence or absence of AICAR (1 mM). These cells were stained with a Pdlim5 antibody, phalloidin and anti-paxillin antibody. Magnified images outlined by yellow squares show the areas outlined by white squares. Scale bar, 10 μm.
