Fig. 3

Association between CD44 and Epac1. A Representative examples and mean ± SE analysis for co-immunoprecipitation, reverse co-immunoprecipitation and cell input of CD44 and Epac1, Epac2, PAKIIα and PKAc in HL-1 myocytes treated with osteopontin, isoproterenol or 8-CPT-cAMP. B Representative examples and mean ± SE analysis for co-immunoprecipitation and cell input of His-tagged CD44 and Epac1 in HL-1 myocytes treated with 8-CPT-cAMP. In A and B the mean ± SE analysis was for precipitated protein-bound Epac1, Epac2, PAKIIα or PKAc or CD44 to precipitated protein ratio and cell input, both of which were quantified to GADPH and normalized to the control. ISO = isoproterenol. The picture is a representation of blots from 3 independent experiments. In A and B, n = 3 per group. C Representative examples and mean ± SE confocal images of proximity ligation assay in HL-1 myocytes. The red color (arrows) indicated the linkage between CD44 and Epac1. The blue color indicated nuclei. The relative fluorescence was normalized to control as 1.0. n = 4 images per group. For A–C, *P < 0.05 versus control by one-way ANOVA with Bonferroni’s post hoc test. ISO = isoproterenol