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PKG1 Is Necessary for Myofilament-Specific β 1 AR Signaling to Promote Protein Phosphorylation and Cardiac Contractility (A, B) WT mice hearts were perfused with saline (Ctrl), ISO (0.1 μmol/L), DOB (1 μmol/L), or CAR (1 μmol/L) for 10 minutes, and the phosphorylation of vasodilator-stimulated phosphoprotein (VASP) at serine 157 and <t>serine</t> <t>239</t> was probed and quantified (n = 5). (C, D) PKG1-flox/flox (PKG1-FF), cardiac deletion of PKG1 (PKG1-CKO), CRE, WT, and whole-body deletion of PKG2 (PKG2-KO) mice were subjected to echocardiographic measurements before and after stimulation with CAR (intraperitoneal injection, 100 μg/kg). Cardiac EF was quantified in PKG1-flox, PKG1-CKO, and CRE mice (n = 8) (C) and in PKG2-KO and WT mice (n = 8) (D). (E, F) PKG1-FF and PKG1-CKO hearts were subjected to Langendorf perfusion with saline and CAR (1 μmol/L) for 10 minutes. Heart tissues were lysed to probe the phosphorylation of phospholamban (PLB), MYPT1, and MLC (n = 3). Data are presented as mean ± SEM. ∗∗ P < 0.01, ∗∗∗ P < 0.01, and ∗∗∗ P < 0.001 compared with basal condition or between indicated groups. P values were calculated using 1-way analysis of variance with Tukey’s post hoc or paired Student’s t -tests. Abbreviations as in <xref ref-type=Figure 1 , Figure 2 , Figure 3 . " width="250" height="auto" />
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PKG1 Is Necessary for Myofilament-Specific β 1 AR Signaling to Promote Protein Phosphorylation and Cardiac Contractility (A, B) WT mice hearts were perfused with saline (Ctrl), ISO (0.1 μmol/L), DOB (1 μmol/L), or CAR (1 μmol/L) for 10 minutes, and the phosphorylation of vasodilator-stimulated phosphoprotein (VASP) at serine 157 and serine 239 was probed and quantified (n = 5). (C, D) PKG1-flox/flox (PKG1-FF), cardiac deletion of PKG1 (PKG1-CKO), CRE, WT, and whole-body deletion of PKG2 (PKG2-KO) mice were subjected to echocardiographic measurements before and after stimulation with CAR (intraperitoneal injection, 100 μg/kg). Cardiac EF was quantified in PKG1-flox, PKG1-CKO, and CRE mice (n = 8) (C) and in PKG2-KO and WT mice (n = 8) (D). (E, F) PKG1-FF and PKG1-CKO hearts were subjected to Langendorf perfusion with saline and CAR (1 μmol/L) for 10 minutes. Heart tissues were lysed to probe the phosphorylation of phospholamban (PLB), MYPT1, and MLC (n = 3). Data are presented as mean ± SEM. ∗∗ P < 0.01, ∗∗∗ P < 0.01, and ∗∗∗ P < 0.001 compared with basal condition or between indicated groups. P values were calculated using 1-way analysis of variance with Tukey’s post hoc or paired Student’s t -tests. Abbreviations as in <xref ref-type=Figure 1 , Figure 2 , Figure 3 . " width="100%" height="100%">

Journal: JACC: Basic to Translational Science

Article Title: Carvedilol Activates a Myofilament Signaling Circuitry to Restore Cardiac Contractility in Heart Failure

doi: 10.1016/j.jacbts.2024.03.007

Figure Lengend Snippet: PKG1 Is Necessary for Myofilament-Specific β 1 AR Signaling to Promote Protein Phosphorylation and Cardiac Contractility (A, B) WT mice hearts were perfused with saline (Ctrl), ISO (0.1 μmol/L), DOB (1 μmol/L), or CAR (1 μmol/L) for 10 minutes, and the phosphorylation of vasodilator-stimulated phosphoprotein (VASP) at serine 157 and serine 239 was probed and quantified (n = 5). (C, D) PKG1-flox/flox (PKG1-FF), cardiac deletion of PKG1 (PKG1-CKO), CRE, WT, and whole-body deletion of PKG2 (PKG2-KO) mice were subjected to echocardiographic measurements before and after stimulation with CAR (intraperitoneal injection, 100 μg/kg). Cardiac EF was quantified in PKG1-flox, PKG1-CKO, and CRE mice (n = 8) (C) and in PKG2-KO and WT mice (n = 8) (D). (E, F) PKG1-FF and PKG1-CKO hearts were subjected to Langendorf perfusion with saline and CAR (1 μmol/L) for 10 minutes. Heart tissues were lysed to probe the phosphorylation of phospholamban (PLB), MYPT1, and MLC (n = 3). Data are presented as mean ± SEM. ∗∗ P < 0.01, ∗∗∗ P < 0.01, and ∗∗∗ P < 0.001 compared with basal condition or between indicated groups. P values were calculated using 1-way analysis of variance with Tukey’s post hoc or paired Student’s t -tests. Abbreviations as in Figure 1 , Figure 2 , Figure 3 .

Article Snippet: Protein supernatants were solved on sodium dodecyl sulfate polyacrylamide gel electrophoresis gels and detected with anti-NOS3 (1:1,000; 9572, Cell Signaling Technology), anti-phospho-NOS3 serine 1177 (1:1,000; 9571, Cell Signaling Technology), anti-β 1 AR (1:500; SC-568, Santa Cruz Biotechnology), anti–vasodilator-stimulated phosphoprotein (1:500; SC-46668, Santa Cruz Biotechnology), anti–phospho–vasodilator-stimulated phosphoprotein serine 157 (1:500; 3111, Cell Signaling Technology), anti–phospho–vasodilator-stimulated phosphoprotein serine 239 (1:500; 3114, Cell Signaling Technology), anti-phospho-phospholamban serine 16 (1:1,000; 14388.

Techniques: Saline, Injection