Fig 2: Enolase 1 (ENO1) is upregulated in stem cell-like cells enriched from PAMC-82 and SNU16 cells by spheroid-forming culture.A Analysis of the self-renewal abilities of PAMC-82, SNU16 parental cells, and spheroids using methylcellulose spheroid-formation assay. Scale bar, 100 µm. B Tumorigenicity assay in PAMC-82, SNU16 parental cells, and spheroids. C Western blot analysis for the expression of ENO1 in PAMC-82, SNU16 parental cells, and spheroids. spheroids: GSCSs enriched from parental cells cultured in spheroid-formation conditions and passaged to the third-passage. Results are from representative experiments in triplicate and shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 3: NDUFA4 promotes glycolytic and oxidative metabolism in GC cells.A ECAR, (B) OCR, (C) lactate, (D) ATP content and (E) expression of ENO1 and LDHA in AGS, HGC27 and MKN45 cells with or without NDUFA4 overexpression or knockdown. ***P < 0.001 vs shNC or vector.

Fig 4: Enolase 1 (ENO1) increases the stemness of gastric cancer (GC) cells via glycolysis promotion.A PAMC-82 and SNU16 cells stably expressing pLenti-NC, pLenti-ENO1, shcon, or shENO1 were cultured for 36 h, then the levels of glucose consumption and lactic acid production were measured according to the cell numbers. Fold changes were normalized (µmol/106 cells). B Extracellular acid ratio (ECAR) was measured by Seahorse XF in PAMC-82 and SNU16 cells stably expressing pLenti-NC, pLenti-ENO1, shcon, or shENO1. ECAR curves from cells treated with glucose, oligomycin, and 2-DG. Black arrows indicate the time point of cell treatment. Results are from representative experiments in triplicate and shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 5: Proteome-wide differences in steady-state protein levels between mouse and naked mole rat cells.A, the volcano plot of the p-value versus log2 ratio of expression levels in naked mole rat and mouse cells. Blue points represent all proteins, and red points highlight proteins involved in glycolysis and oxidative phosphorylation with significantly altered expression levels. B, distribution of log2 expression level ratios for specified protein subsets. The box plot representations are as described in Figure 1. Red boxes highlight GO terms involved in ATP production. *, and *** indicate p-values of less than 0.05, 0.005, and 0.0005, respectively, in comparison with the global distribution using the Mann–Whitney U test. C, western blots of selected proteins from respiratory chain complexes (Uqcrc1, Atp512, Cox4) and glycolysis (Eno1, Gapdh). D, measurements of geometric mean intensities of MitoTracker mitochondrial staining of naked mole rat and mouse cells. GO, gene ontology.