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. 2024 Jan 11;43(3):362–390. doi: 10.1038/s44318-023-00009-w

Figure 8. SQSTM1 mediates NNMT transcription and inhibition of NNMT ameliorates cardiac dysfunction in cAtg3-KO mice.

Figure 8

(A) Protein levels of ATG3, SQSTM1, NNMT, RELA (NF-κB p65) and RELA phosphorylation at S536, and GAPDH in H9c2 cells. ATG3 and SQSTM1 were silenced as indicated, n = 4–8 per group. Data are mean ± SEM. One-way ANOVA followed by Bonferroni’s multiple comparison tests was used to determine statistical significance. *P < 0.05. (B) NAD+ levels in H9c2 cells. Atg3 and SQSTM1 were silenced as indicated, n = 3–4 to 8 per group. Data are mean ± SEM. One-way ANOVA followed by Bonferroni’s multiple comparison tests was used to determine statistical significance. *P < 0.05. (C) Oxygen consumption rate (OCR) in H9c2. ATG3 and SQSTM1 were silenced as indicated, n = 9–30 per group. Data are mean ± SEM. Unpaired t tests were used to determine statistical significance between two groups at corresponding time points. *P < 0.05 vs. Con si, #P < 0.05 vs. Atg3 si. (D) Protein levels of GAPDH, SQSTM1, NNMT, RELA phosphorylation at S536, and RELA in cardiac-specific SQSTM1 KO mouse hearts, n = 3–6 per group. Data are Mean ± SEM. Unpaired t tests were used to determine statistical significance between two groups. *P < 0.05. (E) NAD+ levels in cardiac-specific SQSTM1 KO mouse hearts. n = 3–6 per group. Data are mean ± SEM. An unpaired t test was used to determine statistical significance between two groups. *P < 0.05. (F) Protein levels of SQSTM1, LC3 (MAP1LC3A), and GAPDH in WT and cAtg3-KO mouse hearts. Mice, at ~30 weeks of age, were injected with 5-Amino-1-methylquinolinium iodide (NNMTi) or Veh as indicated. n = 3 per group. Typical blots are shown. (G) NAD+ levels in WT and cAtg3-KO mouse hearts. Mice, at ~30 weeks of age, was injected with NNMTi or Veh as indicated. n = 3–8 per group. Data are mean ± SEM. One-way ANOVA followed by Bonferroni’s multiple comparison tests was used to determine statistical significance. *P < 0.05. (H) Heart rates (HR) of WT and cAtg3-KO mice under isoflurane anesthesia. Mice, at ~30 weeks of age, were injected with NNMTi or Veh as indicated. n = 5–8 per group. (I) Fractional shortening (FS) in WT and cAtg3-KO mouse hearts under isoflurane anesthesia. Mice, at ~30 weeks of age, were injected with NNMTi or Veh as indicated. n = 5–8 per group. Data are mean ± SEM. One-way ANOVA followed by Bonferroni’s multiple comparison tests was used to determine statistical significance. *P < 0.05. Source data are available online for this figure.