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Fig. 3 | Cardiovascular Diabetology

Fig. 3

From: MiR-30c/PGC-1β protects against diabetic cardiomyopathy via PPARα

Fig. 3

PGC-1β expression was reduced by miR-30c. a Venn diagram showing the overlap number of microRNA targeting PGC-1β predicted by TargetScan, PicTar and TarBase websites. b Sequence alignment of miR-30c on the 3′ UTR of PGC-1β from different organisms. c miR-30c expression was determined by RT-PCR in the heart tissues from 28-week old male db/db diabetic mice and their C57 littermates (n = 3). d miR-30c expression was determined by RT-PCR in H9c2 cells with or without palmitate (1 mmol/L) treatment. e Animals (db/db mice and C57 control mice) were injected with the corresponding rAAVs at 8–10 weeks of age and sacrificed at 28 weeks of age (n = 8–10). f H9c2 cells were transfected with miR-30c mimics/inhibitors (or their control) and then subjected to palmitate (1 mmol/L) stimulation. Representative Western blots and quantitative analysis of PGC-1β in the cardiac tissues (e) and H9c2 cells (f). β-actin was used as an internal control. g PGC-1β mRNA level was determined by RT-PCR in the whole RNA and the RNA of the anti-ago2 co-IP complex from H9c2 cells treated with miR-30c mimics or miR-control. h Schematic diagram of luciferase reporter plasmids pMIR PGC-1β 3′ UTR and mutant PGC-1β 3′ UTR, and the potential (or mutant) seed sequence of miR-30c targeting PGC-1β 3′ UTR are shown in red. i Regulation of PGC-1β via miR-30c targeting its 3′ UTR was determined with luciferase reporter assays in HEK293T cells. For d, f, g and i, n = three independent experiments. Data are expressed as mean ± SEM, *p < 0.05. UTR untranslated region, RT-PCR real-time polymerase chain reaction, rAAV recombinant adeno-associated virus, IP immunoprecipitation

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