Extended Data Fig. 3: Effects of insulin treatment on senescent cells in visceral adipose tissue.
From: SGLT2 inhibition eliminates senescent cells and alleviates pathological aging
a, Protocol of the experiments to test the senolytic effects of short-term treatment with insulin. NC, normal chow; HFD, high-fat diet. b, Body weight and gonadal white adipose tissue (gWAT) weight of mice fed NC or HFD with or without insulin (Ins) (n = 12 each). c, Glucose tolerance test and insulin tolerance test of mice as prepared in Extended Data Fig. 3b (n = 6 each). d, SA-β-gal activity (n = 12 each) and hematoxylin and eosin (HE) staining (n = 6 each) in gWAT of mice as prepared in Extended Data Fig. 3b. e, Western blot analysis for p53 in gWAT of mice as prepared in Extended Data Fig. 3b (n = 12 each from 4 gels/blots processed in parallel). f, qPCR analysis for Cdkn1a and Cdkn2a in gWAT of mice as prepared in Extended Data Fig. 3b (n = 12 each). g, qPCR analysis for Ccl2 and Tnf in gWAT of mice as prepared in Extended Data Fig. 3b (n = 12 each). Data were analyzed by two-way ANOVA followed by Tukey’s multiple comparison test (for equal variance) or Dunnett’s multiple comparison test (for unequal variance) (b, d–g), repeated measures analysis (c), or two-tailed unpaired Student’s t-test (c). *P < 0.05, **P < 0.01. Exact P-value: NC versus HFD: < 0.0001 (Body weight and gWAT weight) and 0.0011 (Fasting blood glucose), NC versus HFD+Ins: < 0.0001 (Body weight and gWAT weight), HFD versus HFD+Ins: 0.0001 (Fasting blood glucose) (b); HFD versus HFD+Ins: < 0.0001 (GTT-trend and ITT-trend), 0.0029 (GTT-AUC) and 0.0044 (ITT-AUC) (c); NC versus HFD: 0.0001 (SA-β-gal activity) and 0.0262 (Crown-like structure count), NC versus HFD+Ins: 0.0003 (SA-β-gal activity) and 0.0022 (Crown-like structure count) (d); NC versus HFD: 0.0152, NC versus HFD+Ins: 0.033 (e); NC versus HFD: 0.0092 (Cdkn1a) and 0.0003 (Cdkn2a), NC versus HFD+Ins: 0.0001 (Cdkn1a) and < 0.0001 (Cdkn2a) (f); NC versus HFD: < 0.0001 (Ccl2 and Tnf), and NC versus HFD+Ins: < 0.0001 (Ccl2 and Tnf) (g). Data are shown as the mean ± SE in plots of all individual data (b–g) or as the mean ± SE in the spaghetti plot shown in Supplementary Fig. 1 (c).