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. 2023 Mar 8;132(7):828–848. doi: 10.1161/CIRCRESAHA.122.321448

Figure 4.

Figure 4.

PDE3A (phosphodiesterase 3A) isoforms are in a nuclear complex involving SMAD4 (SMAD family member 4) and HDAC-1 (histone deacetylase 1). A, Western blot analysis showing PDE3A2 and HDAC-1 in the pull down of endogenous SMAD4 obtained from neonatal rat ventricular myocyte (NRVM) cell lysates. Representative of n=3 independent cultures. B, Detection of endogenous SMAD4 and HDAC-1 in the immunoprecipitate obtained by pulling down endogenous PDE3A from NRVM lysates. n=3. C, Immunostaining of endogenous PDE3A and SMAD4 in NRVM cells treated either with DMSO (control) or with leptomycin B (LMB; 100 nmol/L) for 3 hours. Quantification of fluorescence intensity is shown on the right. n=4 independent experiments (at least 15 cells per experiment). Mann-Whitney U test. D, Western blot analysis showing endogenous PDE3A1 and PDE3A2 in the nuclear (N) and cytoplasmic (C) fractions obtained from NRVM treated with 100 nmol/L LMB or dimethyl sulfoxide (DMSO) (control) for 3 hours. Densitometric quantification is shown on the right. Values are shown as relative to the cytosolic content and are presented as mean±SEM. n=6 biological replicates. Wilcoxon matched pairs signed-rank test. WCL indicates whole-cell lysate.