Fig 1: KLF9 knockdown suppresses cardiac dysfunction in DCM. (A) The protein level of KLF9 in mice receiving AAV9-shKLF9 injection (n = 6). (B) Immunofluorescence staining of KLF9 and α-actin in mouse hearts 6 weeks after AAV9-shKLF9 injection (n = 5). (C) Blood glucose in DCM mice at the indicated time points (n = 12). (D) The body weight of DCM mice at the indicated time points (n = 12). (E) H&E staining and cross-sectional area results (n = 6). (F) PSR staining and collagen volume results (n = 6). (G) The mRNA levels of hypertrophy and fibrosis markers in each group (n = 6). * p < 0.05.
Fig 2: KLF9 overexpression aggravates cardiac dysfunction in DCM: (A) The protein level of KLF9 in mice 6 weeks after AAV9-KLF9 injection (n = 6). * p < 0.05 vs. KLF9 0 w. (B) Immunofluorescence staining of KLF9 and α-actin in mouse hearts 6 weeks after AAV9-KLF9 injection (n = 5). (C) Blood glucose in DCM mice at the indicated time points (n = 12). (D) The body weight of DCM mice at the indicated time points (n = 12). (E) H&E staining and cross-sectional area results (n = 6). (F) PSR staining and collagen volume results (n = 6). (G) The mRNA levels of hypertrophy and fibrosis markers in each group (n = 6). * p < 0.05.
Fig 3: Expression of KLF9 is increased by Dex in RAW264.7 cells. (A) Real-time quantitative PCR analysis of the mRNA level of KLF9 in RAW264.7 cells 16 h after treatment with saline or Dex (100 nM) (n=5). (B) Western blot analysis of KLF9 in RAW264.7 cells as described in A; quantitative data are on the right. (C) A series of promoter activity of KLF9 was detected in RAW264.7 cells with saline or Dex (100 nM) treatment. (D) A ChIP assay was performed using anti-GR antibody in RAW264.7 cells. (E) Real-time quantitative PCR analysis of KLF9 in RAW264.7 cells treated with 100 nM Dex or with 10 µM of the GR antagonist RU486 for 16 h. (F) Western blot analysis of the protein level of KLF9 described in E; quantitative data are on the right. Experiments were performed in triplicate. Error bars represent the standard deviation *P<0.05; **P<0.01; ***P<0.005; ****P<0.001. KLF9, Krüppel-like factor 9; Dex, dexamethasone; ChIP, chromatin immunoprecipitation; GR, glucocorticoid receptor.
Fig 4: KLF9 knockdown ameliorates inflammation and oxidative stress in DCM. (A) CD68 staining and quantitative results in DCM mice receiving AAV9-shKLF9 injection (n = 5). (B) Cytokine levels in heart tissue in DCM mice receiving AAV9-shKLF9 injection (n = 6). (C) DHE staining in DCM mice receiving AAV9-shKLF9 injection (n = 5). (D) MDA levels and NADPH oxidase activity in DCM mice receiving AAV9-shKLF9 injection (n = 6). (E) SOD2 and Gpx activity (n = 6). * p < 0.05.
Fig 5: PPARγ/NRF2 inhibition counteracted the protective effects of KLF9 knockdown. (A–D) Cardiomyocytes were transfected with KLF9 siRNA, treated with the PPARγ inhibitor GW9662 (10 μM), or an NRF2 inhibitor (ML385, 1 μM) for 12 h and were then exposed to HG for 48 h. (A) Cytokine levels in cells, as detected by ELISA (n = 6). (B) ROS levels (n = 6). (C) MDA levels and NADPH oxidase activity (n = 6). (D) SOD2 and Gpx activity in cells (n = 6). * p < 0.05.
Supplier Page from ABclonal Technology for KLF9 Rabbit pAb
Trial Size: 20 ul