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. 2016 Nov 11;6:36803. doi: 10.1038/srep36803

Figure 2. MiR-208a suppresses PDE4D in adult ventricular myocytes.

Figure 2

(A) Two sites of PDE4D 3′ UTR sequence alignment are shown across species highlighting the seed match sequences of PDE4D 3′UTR (red) complementary to the seed sequences of miR-208a. Expression of PDE4D as assessed by immunohistochemistry (B–D). Significant reduction of total PDE4D staining was observed using a PDE4D specific antibody both in the cytoplasm and nucleus of miR-208a transduced but not in control myocytes (B–D). Sarcomeric actinin (green) shows the presence of the myocytes that were also stained by PDE4D (D; small box green for actinin). The images were taken with 40x objective confocal microscopy and bar shows scale of 50 μm. Western blots show that PDE4D was reduced significantly in miR-208a transduced but not in untreated or miR-208a mutant myocytes (E,F). Effects of pharmacologic inhibition of miR-208a via miR-208a power inhibitor that increased PDE4D content (G,H). Note that the Western blot in G was run longer resulting in distinct band separation versus E. Acute suppression of miR-208a in adult ventricular cardiac myocytes by antimiR-208a (I,J). Data are shown as mean +/− SEM, n = 3 *P < 0.05.