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. 2024 Aug 26;15(5):2104–2117. doi: 10.1002/jcsm.13558

Figure 4.

Figure 4

Betaine attenuated Yy1 age‐related decline and Yy1 transcriptionally downregulated the Mss51 transcription. (A) Six potential candidates were figured out by utilizing three databases. Yy1 mRNA expressions were significantly higher in YOU and BET group. Mean ± SD, n = 4/group. (B) WB revealed that Yy1 protein level declined with aging and could be preserved by betaine treatment. Mean ± SD, n = 4/group. (C) GEPIA2 database indicates a negative correlation between the expression Yy1 and Mss51. (D) The full‐length promoter (T‐2000) and a series of truncated promoter (T‐1000, T‐350, and T‐200) into pGL4.10 luciferase reporter plasmids were co‐transfected with Yy1 plasmid in C2C12 cells. Relative fluorescence intensity (RFI) from Luciferase assay was compared, showing the downregulation of Yy1 on Mss51 transcription. aSignificant different RFI compared with vector group. bSignificant difference by post‐hoc analysis for comparison among T‐2000, T‐1000, T‐350, and T‐200 groups. c Significant different RFI between T‐350 and T‐200 groups. Mean ± SD, n = 4/group. (E) ChIP results demonstrated that Yy1 could bind to the region (−350 bp to +100 bp) of the Mss51 promoter. Mean ± SD, n = 4/group. (F) EMSA showed that Yy1 could specifically bind to the Mss51 promoter region with the sequence (5′ … CAAAATGG … 3′). *P < 0.05; **P < 0.01; ***P < 0.001.