Fig 1: The expression levels of CTRP13 in H9c2 cells with or without H/R stimulation. The mRNA and protein levels of CTRP13 were detected using qRT-PCR and western blot, respectively. (A) Results from qRT-PCR. (B) Results from western blot. n = 4. *P < 0.05 vs. control group.
Fig 2: Recombinant CTRP13 protein attenuated myocardial I/R-induced injury in rats. (A) The transduction efficiency of CTRP13 in the I/R rats. (B, C) Illustration of HE staining and TTC staining of the myocardial tissues in different rat groups. (C) Myocardial infarct size was determined. (D) Representative photomicrographs (×200) of TUNEL staining in myocardial I/R tissue. Green fluorescence indicates TUNEL-positive apoptotic nuclei; blue fluorescence indicates total cardiomyocyte nuclei. (E) ROS were stained with DHE (red) and nuclei with DAPI (blue). (F, G) The serum levels of LDH and CK were evaluated using commercial ELISA kits. n = 5. *P < 0.05 vs. Sham group; #P < 0.05 vs. I/R group.
Fig 3: Effect of AMPK inhibition on CTRP13-mediated activation of Nrf2/ARE signaling pathway in H/R-induced H9c2 cells. H9c2 cells were transfected with pcDNA3.1-CTRP13 for 48 h in the presence of Compound C (10 μM), and then they were subjected to H/R injury. (A) Nrf2 nuclear protein expression was measured by western blot. (B) Quantification analysis of Nrf2. (C) Luciferase reporter assay was conducted to explore the Nrf2 transcriptional activity. (D) Cell viability in H9c2 cells. (E) ROS production in H9c2 cells. (F) Caspase-3 activity in H9c2 cells. n = 4. *P < 0.05 vs. control group; #P < 0.05 vs. H/R+pcDNA3.1 group; &P < 0.05 vs. H/R+pcDNA3.1-CTRP13 group.
Fig 4: Overexpression of CTRP13 improved the viability of H9c2 cells exposed to H/R. H9c2 cells were transfected with pcDNA3.1-CTRP13 or pcDNA3.1 for 48 h, then they were exposed to H/R injury. (A and B) The efficiency of transfection was evaluated using qRT-PCR and western blot. (C) Cell viability was measured by MTT assay. n = 4. *P < 0.05 vs. control group; #P < 0.05 vs. H/R+pcDNA3.1 group.
Fig 5: Overexpression of CTRP13 enhanced the activation of AMPK/Nrf2/ARE signaling pathway in H/R-stimulated H9c2 cells. (A-C) Western blot was performed to detect the expression levels of AMPK, p-AMPK and nuclear Nrf2. (D) Luciferase reporter assay was conducted to explore the Nrf2 transcriptional activity. n = 3. *P < 0.05 vs. control group; #P < 0.05 vs. H/R+pcDNA3.1 group.
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