Fig. 5: Overexpression of Psmb8 reduces H/R-stimulated cardiomyocyte apoptosis, ROS level and mitochondrial fission in vitro. | Cell Death & Disease

Fig. 5: Overexpression of Psmb8 reduces H/R-stimulated cardiomyocyte apoptosis, ROS level and mitochondrial fission in vitro.

From: Psmb8 inhibits mitochondrial fission and alleviates myocardial ischaemia/reperfusion injury by targeting Drp1 degradation

Fig. 5

A Neonatal rat cardiomyocytes (NRCMs) were infected with adenovirus expressing Ad-GFP or Ad-Psmb8 for 24 h and then simulated with H/R for another 24 h. Immunostaining of NRCMs with TUNEL (red) and DAPI (blue) to detect apoptotic cardiomyocytes (left). Quantification of the percentages of TUNEL-positive nuclei (right, n = 3 independent experiments). Bar: 50 μm. B Immunostaining of mitochondrial morphology was performed in cardiomyocytes using MitoTracker® Red (left). Quantification of fragmented mitochondria (right, n = 3 independent experiments). Bar: 10 μm. C NRCMs were infected with adenovirus expressing siRNA-control or siRNA-Psmb8 for 24 h and then stimulated with H/R for another 24 h. Immunostaining of NRCMs with TUNEL (red) and DAPI (blue) was performed to detect apoptotic cardiomyocytes (left). Quantification of the percentages of TUNEL-positive nuclei (right, n = 3 independent experiments). Bar: 50 μm. D Immunostaining of mitochondrial morphology in cardiomyocytes with MitoTracker® Red (left) and analysis of fragmented mitochondria (right, n = 3 independent experiments). Bar: 10 μm. E Detection of the Psmb8, Drp1, p-Drp1(S616), Mfn1, and Mfn2 protein levels with immunoblotting (left) and analysis of each protein level (right, n = 4 each group). F Detection of the Psmb8, Drp1, p-Drp1 (S616), Mfn1, Mfn2, Mito-Drp1 and Mito-Drp1 (S616) protein levels with immunoblotting (left) and analysis of each protein level (right, n = 4 each group). All values are expressed as the mean ± SEM, and n indicates the sample numbers per group.

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