Fig. 6: Increased PPARGC1 and ESRRA alleviate mitochondrial and cardiac dysfunction due to Hey2 overexpression. | Nature Communications

Fig. 6: Increased PPARGC1 and ESRRA alleviate mitochondrial and cardiac dysfunction due to Hey2 overexpression.

From: The transcriptional repressor HEY2 regulates mitochondrial oxidative respiration to maintain cardiac homeostasis

Fig. 6

a MitoTracker Red CMXRos staining measuring the mitochondrial mass of control AC16 or HEY2-OE AC16 cells transfected with control vectors, PPARGC1A or ESRRA vectors, as well as PPARGC1A and ESRRA plasmids together. Scale bar: 50 µm. b Quantification of MitoTracker Red fluorescent intensity of transfected HEY2-OE AC16 cells and control AC16 cells. Data were relative to fluorescent intensity in control group. n = 38 (ctrl+vector, OE + PPARGC1A + ESRRA), n = 41 (ctrl+PPARGC1A), n = 39 (ctrl+ESRRA), n = 43 (the rest groups). Box plots displayed median ± interquartile range (IQR), with whiskers extending to the minimum and maximum values. *P < 0.05, **P < 0.01, ****P < 0.0001, Student’s t-test (unpaired, two-tailed). c ATP levels of control AC16 and HEY2-OE AC16 cells transfected with control vectors, PPARGC1A or ESRRA vectors, and PPARGC1A and ESRRA plasmids together. Data were relative to ATP level in control group. 3 biological replicates. d RT-qPCR analysis showing ETC complex genes that expressed in control AC16 and HEY2-OE AC16 cells transfected with control vectors, PPARGC1A or ESRRA vectors, and PPARGC1A and ESRRA plasmids together. 3 biological replicates. e Measurement of mitochondrial ETC complex I activity in control AC16 and HEY2-OE AC16 cells transfected with control vectors, and PPARGC1A and ESRRA plasmids together. 3 biological replicates. f Schematic synthesis diagram of Flag-ppargc1a and HA-esrra mRNA with N1mΨ and Cap analogs modifications. g Experimental diagram for 4-HT treatment, modRNAs injection and cardiac analysis. h Echocardiography analyses depicting EF and FS in hey2-mOE and control hearts injected with vehicle, ppargc1a modRNA, esrra modRNA, or ppargc1a and esrra modRNAs together. n = 10 (hey2-mOE+ppargc1a+esrra), n = 11 (the rest groups). i RT-qPCR analysis showing ETC complex genes that differentially expressed in hey2-mOE and control hearts injected with vehicle, ppargc1a modRNA, esrra modRNA, or ppargc1a and esrra modRNAs together. 3 biological replicates. Data presents as mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, Student’s t-test (unpaired, two-tailed) (c, e, h) or Multiple t tests (two-tailed) (d, i). Source data are provided as a Source Data file.

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