Journal: Signal Transduction and Targeted Therapy
Article Title: Abrogation of aberrant glycolytic interactions eliminates senescent cells and alleviates aging-related dysfunctions
doi: 10.1038/s41392-025-02502-6
Figure Lengend Snippet: A PGAM1-Chk1 antagonist suppresses FOXM1 during senescence. a , b Assessment of the proapoptotic BH family in SnCs after Nutlin 3b treatment. a RT‒PCR analysis of SnCs with or without Nutlin 3b. ( n = 3, biological replicates). b p53 siRNA-transfected SnCs were exposed to Nutlin 3b. RT‒PCR analysis of the indicated cells. c Knockdown of BIM in SnCs. SnCs were transfected with the indicated siRNA, siBIM, or scramble control RNA. The transfectants were then subjected to Nutlin 3b treatment. d – f Comparison of the results of the microarray analysis between the control and Nutlin 3b-treated SnCs ( n = 3, biological replicates). d Fold enrichment analysis by comparison between 1168 genes downregulated by Nutlin 3b in SnCs and 9442 datasets of chromatin immunoprecipitation (ChIP) information reported in the National Center for Biotechnology Information (NCBI), European Bioinformatics Institute (EBI) and DNA Data Bank of Japan (DDBJ). Eighty-six of the 9442 datasets displayed high enrichment (>3-fold). Twenty-one datasets for FOXM1 are shown in red (left panel). e Heatmap of 37 FOXM1 target genes downregulated by Nutlin 3b. SnCs with or without Nutlin 3b were compared. f Gene expression of the FOX family in SnCs after Nutlin 3b treatment, as determined via microarray data. Left panel, heatmap analysis of the FOX family. Right panel: Red and blue bars indicate reduced and increased expression, respectively. Dagger (†) indicates |FC|≥2 and Lpe. P < 0.05 is presented in Supplementary Fig. . Evaluation of FOXM1 protein levels in SnCs after Nutlin 3b treatment ( g ) and after transfection with HIF-2α siRNA ( h ). The data are representative of two independent experiments. i Evaluation of BIM mRNA in SnCs after FOXM1 knockdown. FOXM1 siRNA was transfected into SnCs ( n = 3, biological replicates). j Schematic diagram of the promoter region in the human BIM gene. Several cis-elements for FKH are shown. The indicated regions were subsequently cloned and inserted into luciferase reporter plasmids (left lower panel). Luciferase reporter assays were performed in SnCs after Nutlin 3b treatment (middle panel). A reporter assay for the proximal region was conducted in SnCs after FOXM1 knockdown (right panel) ( n = 3, biological replicates). The data represent the means ± SEMs. Single (*) and double (**) asterisks indicate statistical significance at p < 0.05 and p < 0.01, respectively. Statistical analyses were performed via unpaired Student’s two-tailed t test ( a ) or one-way analysis of variance (ANOVA) and Dunnett’s multiple comparison test ( b , c , i and j )
Article Snippet: Anti-mouse Chk1 (25887-1-AP, 1:1000) and anti-mouse FOXM1 (13147-1-AP, 1:1000) were purchased from Proteintech (USA).
Techniques: Transfection, Knockdown, Control, Comparison, Microarray, Chromatin Immunoprecipitation, Gene Expression, Expressing, Clone Assay, Luciferase, Reporter Assay, Two Tailed Test