Fig. 8: ELF1-mediated transcriptional regulation of YTHDF2 promotes NPC senescence.

A The transcription factor database (http://www.grndb.com/) for single-cell sequencing was used to predict the potential transcription factors regulating YTHDF2 expression. B, C Western blot and qPCR analyses were performed to detect the Ythdf2 protein and mRNA expression levels in NP tissues after Elf1 knockout (KO). D The YTHDF2 protein expression levels were detected using western blot analysis following ELF1 knockdown in H_NPC. E The Ythdf2 protein expression patterns were detected in NP tissues of Elf1 KO mice using immunofluorescence. F An ELF1 binding peak was detected in the YTHDF2 promoter region using the open ChIP-seq database. G The YTHDF2 promoter sequence was detected by PCR analysis of ELF1 immunoprecipitates. H ELF1 binding to the YTHDF2 promoter in H_NPC was confirmed by ChIP-PCR. I ELF1-like elements in the promoter region of the human YTHDF2 gene. J Luciferase activity, driven by the YTHDF2 promoter, was more pronounced following ELF1 overexpression. In contrast, in the absence of ELF1, the luciferase activity was decreased; The luciferase activity was unchanged with the mutant YTHDF2 reporter following ELF1 overexpression. K The protein expression levels of ELF1, METTL3, and YTHDF2 were detected in human NP tissues using western blot analysis. L, M The Ythdf2 mRNA and protein expression levels were examined using qPCR and immunofluorescence following injection of the Ythdf2-overexpressing lentivirus into mouse NP tissues. N Western blot analysis was used to examine the Ythdf2 protein expression levels in mouse NP tissues. O Grouping of the Ythdf2-overexpressing lentiviruses injected into the NP tissues of Elf KO mice. P, S MRI was used to detect the T2-weighted signal intensity of the intervertebral discs after injection of the Ythdf2-overexpressing lentivirus into the NP tissues of wild-type (WT) and Elf1 KO mice; N = 6. Q, R, T H&E and Safranin-O staining of intervertebral discs after injection of the Ythdf2-overexpressing lentivirus into the NP tissues of WT and Elf1 KO mice; N = 6; Scale bars = 100 μm and 20 μm. 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.