Fig. 6. Silencing RISP mitigates the hyperfunctional RyR2 activity after CH.

a Representative western blot of in vivo specific silencing RISP in PAs rather than liver using shRNA-mediated lentivirus hydrodynamic intravascular (jugular vein) delivery technique. (NS, shRNA of non-silencing; KD, knock down group). b RyR activity in RISP KD group is measured by [³H]-ryanodine binding assays (n = 5 independent studies, 5 mice per study). c Representative western blot of oxidized RyR2 from PAs after exposure to CH in the same film. The oxidation levels are assayed using an anti-2,4-dinitrophenyl (DNP) antibody against ROS-mediated, DNP-derivatized protein carbonyls. d RISP KD suppresses the oxidation of RyR2 and reverses remodeling of FKBP12.6/RyR2 complex determined by western blot. e Silencing RISP in vivo prevents the development of CH-induced PH by measuring RVSP and RV/LV + S (n = 5 independent studies, 5 mice per study). f Silencing RISP inhibits the mitochondrial-mediated and complex III-mediated reactive oxygen species (ROS) generation respectively. Data are expressed as mean ± standard error. (*P < 0.05, using one-way ANOVA test).