Fig. 5

NF-κB signaling pathway supports the long-term maintenance of SG-BPCs. A Volcano plot depicting differential gene expression between KEM and KEM + YAL, as analyzed by bulk RNA sequencing. Data points are colored based on their average expression across all datasets. Red points represent genes upregulated in KEM + YAL, whereas blue points indicate downregulated genes. An adjusted p < 0.1 is considered significant for differential expression. B The gene expression of cellular senescence markers (ADAM28, ID3, TAGLN, and MYB) in SG-BPCs untreated and treated with small molecules. C Western blot of the cells cultured under KEM and KEM + YAL conditions at PD40 for ID3 and β-actin. Full-length blots are presented in Fig. S7C. D Dot plots illustrate the results of GSEA with an adjusted p < 0.1. Each dot plot highlights enriched terms within the transcriptome of KEM + YAL. The size of the dot corresponds to the number of enriched genes, and the color indicates the adjusted p-value. E Western blotting is performed to analyze protein expression levels of p-p65, p65, and β-actin in PD40 cells cultured without or with small-molecule cocktails. Blotting for p-p65, p65, and β-actin are performed sequentially on the same membrane. Full-length blots are presented in Fig. S7D. F Representative images of the cells cultured under KEM, KEM + YAL, and KEM + YAL + NF-κBi conditions for 72 h at PD40 time-point. The data are representative of three independent experiments performed in triplicate and are expressed as mean ± SEM. ns = not significant, * = p < 0.05, ** = p < 0.01