CaM kinase II selectively signals to histone deacetylase 4 during cardiomyocyte hypertrophy
J. Clin. Invest. Johannes Backs, et al. 116:1853 doi:10.1172/JCI27438 [Go to this article.]

Figure 6
Cytosolic accumulation of HDAC4 in cardiomyocytes. (A–C) NRVMs were infected with adenoviral FLAG-HDAC4 or GFP-HDAC5. Subcellular distribution of HDAC4 and HDAC5 was verified following stimulation with PE (20 μM) for 4 hours. NRVMs were pretreated with the kinase inhibitors staurosporine (Stauro; 500 nM), KN93 (5 μM), KN62 (10 μM), AIPII-2 (500 nM), Bis (2.5 μM), Gö6976 (200 nM), or H89 (1 μM). (A) Representative images. (B) Quantitative analysis of time-dependent PE-induced cytosolic accumulation of HDAC4. (C) Effects of kinase inhibitors on PE-induced cytosolic accumulation of HDAC4. (D) NRVMs were treated with PMA with and without Bis. Immunoblotting was performed with antibodies against PKD (lower panel) and phospho-S744/S748 PKD (p-PKD; upper panel). (E–G) NRVMs were infected with adenoviruses encoding FLAG-HDAC4-WT or FLAG-HDAC4-S246,467,632A (FLAG-HDAC4-S/A). One day after infection, cells were grown in serum-free media for 24 hours and then stimulated with PE (20 μM). Cells were fixed and stained with anti-sarcomeric α-actinin (red signal; 24 hours after PE) (E) or anti-ANP (perinuclear green signal; 12 hours after PE) (G). HDAC4-infected NRVMs were identified by anti-FLAG staining (green in E or red in G). (F) [³H]-leucine was added to NRVMs 2 hours after PE stimulation and [³H]-leucine incorporation was measured 24 hours later. *P < 0.05 vs. WT without PE and vs. S/A with PE. NS, not significant vs. S/A without PE. (A, E, and G) Representative images were captured at a magnification of ×40.