Fig. 6: Effects of ENT1/2 modulation on cellular senescence in MSCs.

a, b GSEA plots demonstrated the enrichment of cellular senescence-related genes in MSCs with ENT1 (a) or ENT2 (b) knockdown (n = 3). c, d Micrographs depicting SA-β-gal staining (c) and γH2AX immunofluorescence (d) with ENT1/2 knockdown (n = 3). e, h Heatmaps illustrate DEGs linked to cellular senescence (e) and autophagy (h) in MSCs with ENT1/2 knockdown (n = 3). f, g Western blot images (f) and densitometric analysis (g) of senescence biomarker P16, P21, and P53 (n = 4). i, j Images (i) and quantification (j) of western blot for autophagy markers P62 and LC3B (n = 4). k, l Representative images of SA-β-gal staining (k) and γH2AX immunofluorescence (l) in MSCs overexpressing ENT1/2 with additional treatment of NAM (500 μM). m, n Micrographs showcasing SA-β-gal staining (m) and γH2AX immunofluorescence (n) with ENT1/2 knockdown and supplemented with NMN (250 μM) (n = 3). o Research summary. Scale bars of (c, k, m) represent 100 μm, (d, l, n) represent 50 μm. n represents biologically independent replicates. Data are represented as mean ± s.e.m. P values were calculated using one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons test for (g and j). Gene mRNA and protein expression levels were normalized to β-ACTIN.