Fig. 9: The small-molecule drug mycophenolate mofetil (MMF) targets and interferes with ELF1 expression to inhibit NPC senescence.

A Proteins used for virtual screening of boxes (proteins are surface representations; AI predicted binding pockets are shown in green). B Protein Rasch plot; the number of amino acids located outside the 0.002 curve is small, accounting for 34%, suggesting that this structure can be used for subsequent calculations. C Docking scores for the ELF1 protein. The second to fifth columns of the table represent the Compound Index, Affinity (kcal/mol), Compound Name, and CAS number, respectively. D TOP1 interaction diagram of ELF1. Solid blue line: hydrogen bonding; dashed gray line: hydrophobic interaction. E Detailed view of the 2-dimenstional interaction of ELF1 with top 1 small molecules. F, G Top 1 small molecule structure, with the binding conformation of ELF1 to the small molecules. H MMF IC50 value in H_NPC. I Western blot analysis was used to detect the protein expression levels of ELF1, METTL3, and P21 following MMF treatment in H_NPC. J Western blot analysis was used to detect the protein expression levels of METTL3, MMP13, and E2F3 following MMF treatment in the 10 ng/mL IL-1β model. K–N Immunofluorescence was used to detect the protein expression patterns of ELF1, P21, P16, and Collagen II after the addition of MMF with 48 h of 10 ng/mL IL-1β treatment. O SA-β-gal staining was used to detect the number of senescent NP cells after the addition of MMF with 48 h of 10 ng/mL IL-1β treatment. P SA-β-gal staining was used to detect the amount of H_NPC senescence after the addition of MMF in the replicative senescence model. Q Flowchart of the experiment. Juvenile (2 months old, males, N = 6) or naturally aged (18 months old, males, N = 12) C57BL/6 J mice were given a carrier (carboxymethylcellulose sodium (CMC-Na) or MMF (30 mg/kg/day)) orally. The lumbar vertebrae were collected for histological examination after 6 months of continuous experiments with MRI followed by execution. R, T MRI detection of the T2-weighted signal intensity of the intervertebral discs in MMF-treated mice; N = 6. S, U H&E and Safranin-O staining of mouse intervertebral discs after MMF treatment; N = 6; Scale bars = 100 μm and 20 μm. V Immunofluorescence was used to examine the protein expression patterns of Elf1, p21, and Mettl3, as well as the m6A modifications, in the NP tissues of MMF-treated mice. The data are presented as the mean ± SD. One-way ANOVA was used for comparisons among multiple groups. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.