Figure 3.

Inhibition of PARP1 and Cytosolic Retention of PKM2 Prevent CM Dysfunction Under Exposure to Oxidative Damage

(A) Schematic of the experimental design. (B) Representative immunofluorescent images of PARP1, PKM2, nuclear factor κB (NF-κB), TUNEL, and cytosolic 8-Oxo-2′-deoxyguanosine (8oxodG) in neonatal rat RV CMs (n = 3 or 4) exposed or not to either endothelin-1 (ET-1) or ET-1 + H₂O₂ in the presence or absence of ABT-888, TEPP-46, and DASA-58. (C) Representative western blots (of 3 independent studies) for PARP1, PKM2, NF-κB, α/β tubulin (Tub), and histone H3 in nuclear and cytosolic extracts from H9c2 cells exposed or not to either ET-1 or ET-1 + H₂O₂ in the presence or absence of ABT-888, TEPP-46, DASA-58, and siPARP1. (D) Representative immunofluorescent images of TnT, PKM2, NF-κB, and PARP1 in neonatal CMs isolated from wild-type (Parp1^+/+, n = 3) and Parp1-knockout (Parp1^−/−, n = 3) mice exposed or not to ET-1 + H₂O₂. Scale bars, 25 mm. NT = non-treated; siSCRM = scrambled siRNA; Veh = vehicle; other abbreviations as in Figure 1.