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Fig. 4 | Cardiovascular Diabetology

Fig. 4

From: Acid sphingomyelinase promotes diabetic cardiomyopathy via NADPH oxidase 4 mediated apoptosis

Fig. 4

ASMase knockdown dampens high glucose, palmitate acid and ceramide induced oxidative stress and apoptosis in H9c2 cells. a Representative Western blot images of ASMaseexpression when H9c2 cells were posed to vehicle or pretransfected with siRNA ASMase (si-ASMase), scrambled siRNA (si-control) was used as control (n = 5). bc Representative Western blot and quantitative analysis images of ASMase, NOX2 and NOX4 expression when H9c2 cells were treated with vehicle, glucose + PA, ceramide, siASMase, siASMase + glucose + PA, siASMase + ceramide, respectively. β-actin was used as a loading control. Three to five independent experiments were performed to calculate the means. d Quantitative analysis of NOX1, NOX2 and NOX4 mRNA expression by qPCR in H9c2 cardiomyocytes after transfected by si-ASMase. Four independent experiments were performed to calculate the means. e Representative fluorescence images of rhodamine-phalloidin staining to visualize cell size, scale bar:100 μm. f Quantitative analysis of pro-hypertrophic markers, Nppb and Myh7 mRNA expression by qPCR in H9c2 cardiomyocytes. Four independent experiments were performed to calculate the means. gh Representative fluorescence images and quantitative analysis of DCFH-DA staining for ROS measurement in H9c2 cells, scale bar:100 μm. Three independent experiments were performed to calculate the means. i Representative Western blot images of protein expression of cleaved caspase-3, β-actin was used as a loading control. jk Representative fluorescence images and quantitative analysis of TUNEL staining, scale bar:100 μm. Five independent experiments were performed to calculate the means. Data were presented as mean ± SEM, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

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