Fig 1: GSTM2 attenuates DNA damage in hypertrophic heart tissues. A Immunofluorescence analysis of γ-H2AX (green) in primary cardiomyocytes stimulated with phenylephrine (PE, 50 μmol/l), or DMSO (as a control) for 24 h. Nuclei were stained with DAPI (blue). Scale bar: 50 μm. B Assessment of the 8-OHDG contents in the culture supernatants from PE- or DMSO (as a control)-treated cardiomyocytes. n = 6. *p < 0.05. C Assessment of the 8-OHDG contents in the heart tissues of sham or TAC model mice. n = 6. *p < 0.05. D Representative confocal immunofluorescence images of cardiomyocytes (α-actinin, green) from heart tissues of sham or TAC model mice. The nuclei were stained with DAPI (blue). eccDNA was quantified by analyzing the intensity of DAPI outside nuclei. *p < 0.05. Scale bar: 100 μm. E Assessment of the 8-OHDG contents in clinical human heart failure (HF) tissues and normal heart tissues (as control). n = 3. *p < 0.05. F Representative confocal immunofluorescence images of cardiomyocytes (α-actinin, red) from clinical human HF tissues and normal heart tissues (as control). The nuclei were stained with DAPI (blue). eccDNA was quantified by analyzing the intensity of DAPI outside the nuclei. *p < 0.05. Scale bar: 100 μm. G Immunofluorescence analysis of γ-H2AX (green) in primary cardiomyocytes infected with AAV9-GSTM2 or control AAV9 for 48 h and then treated with PE for 24 h. Scale bar: 200 μm. H Assessment of the 8-OHDG content in the culture supernatants from cardiomyocytes infected with AAV9-GSTM2 or control AAV9 for 48 h and then treated with PE for 24 h. n = 6. *p < 0.05. I Representative confocal immunofluorescence images of cardiomyocytes (α-actinin, green) from the heart tissues of TAC model mice infected with AAV9-GSTM2 or control AAV9. The nuclei were stained with DAPI (blue). eccDNA was quantified by analyzing the intensity of DAPI outside the nuclei. *p < 0.05. Scale bar: 100 μm
Fig 2: Analysis of the differentially expressed genes in samples from heart failure patients. A 2D visualization of GSTT2, GSTZ1 and GSTM2 gene expression in the LV and LA. B Analysis of GSTT2, GSTZ1 and GSTM2 protein abundances according to proteomics. C Immunofluorescence analysis of GSTM2 (red) expression in cardiomyocytes (stained with the cardiomyocyte marker α-actinin, green) from clinical human cardiac hypertrophy (CH) tissues and normal heart tissues. The nuclei were stained with DAPI (blue). Scale bar: 200 μm. D Immunofluorescence analysis of GSTM2 (red) expression in cardiomyocytes (stained with the cardiomyocyte marker α-actinin, green) from the heart tissues of sham or TAC model mice. The nuclei were stained with DAPI (blue). Scale bar: 100 μm. E Representative Masson staining results to assess fibrosis in heart tissues from mice injected with AAV9-GSTM2 or control AAV9 via the tail vein for 3 weeks, then subjected to TAC, and analyzed 4 weeks later. F Statistical analysis of the heart weight (HW)/body weight (BW) ratios of mice subjected to TAC surgery 4 weeks after surgery (n = 6 mice per group; *p < 0.05 versus sham group). G Assessment of left ventricular end-diastolic diameter (LVEDd) in mice, n = 6. *p < 0.05
Fig 3: GSTM2 alleviates IFN-I-stimulated macrophage inflammation. A qRT‒PCR analysis of IL-6 and TNF-α mRNA expression in RAW264.7 macrophages treated with culture supernatants from PE- or DMSO-treated cardiomyocytes. n = 3. *p < 0.05. B qRT‒PCR analysis of IL-6 and TNF-α mRNA expression in the RAW264.7 macrophages treated with culture supernatants from PE-treated cardiomyocytes infected with AAV9-GSTM2 or control AAV9. n = 3. *p < 0.05. C Western blotting analysis of STAT1 and p-STAT1 expression in RAW264.7 macrophages treated with culture supernatants from PE-treated cardiomyocytes infected with AAV9-GSTM2 or control AAV9. GAPDH was used as a loading control. D qRT‒PCR analysis of ISG15, MX1, and CXCL10 mRNA expression in RAW264.7 macrophages treated with culture supernatants from PE-treated cardiomyocytes infected with AAV9-GSTM2 or control AAV9. n = 3. *p < 0.05. E Western blotting analysis of IRF3 and p-IRF3 expression in the cardiomyocytes treated with PE alone or combined with AAV9-GSTM2 infection. GAPDH was used as a loading control. F Assessment of IFN-α and IFN-β levels in the culture supernatants from PE-treated cardiomyocytes infected with AAV9-GSTM2 or control AAV9. n = 3. *p < 0.05. G qRT-PCR analysis of IL-6 and TNF-α mRNA expression in RAW264.7 macrophages treated with culture supernatants from PE- and IFNAR1 antibody-treated cardiomyocytes. n = 3. *p < 0.05
Supplier Page from Abcam for Anti-GSTM2 antibody