Fig. 1: HK expands healthspan and lifespan in mice.
a, Schematic representation of the experimental design. b, Lifespan analysis of treated and untreated mice; n = number of mice tested in each group; the percentage change was calculated with respect to control untreated mice. c, Survival curves for untreated (n = 37) and treated (n = 48) mice. d, Physical appearance score progression in untreated (UT) and treated (T) animals (UT n = 37; T n = 48). e, Quantification of the fur status based on the evaluation score (0–3, where 0 indicates dense fur and 3 indicates thinner fur (UT n = 27; T n = 46; treatment lasted 4 months). f, Representative pictures of 24-month-old mice untreated or HK treated. g, Representative pictures of H&E staining of skin of 24-month-old animals. Insets show enlarged image of dashed box. h, Quantification of hair follicle diameter from sections of 24-month-old untreated or HK-treated mice (n = 6). i,j, Bone tissue percent (i) and bone mineral density (j) of the femurs of young (4 months old) and 24-month-old animals after 4 months of treatment (Bone tissue %, Young n = 6; UT n = 8; T n = 8. Bone mineral density, Young n = 6; UT n = 7; T n = 8). k, Quantification of proteoglycan loss (Young n = 8; UT n = 12; T n = 12). l, OARSI score quantification (Young n = 8; UT n = 10; T n = 12). m, Representative images of Safranin staining for the detection of proteoglycan-rich cartilage of young (4 months old), 24-month-old untreated and 24-month-old treated animals after 4 months of treatment (top images observed at ×10 magnification, bottom (area indicated in dashed box in upper image) at ×40). n, Results of grip strength assay performed on young (3-month) and old (24-month) animals after 4 months of treatment (Young n = 10, UT n = 5, T n = 9). o, Quantification of tibialis muscle fiber CSA (Young n = 7; UT n = 16, T n = 15). p, Representative pictures of tibialis muscle sections with dystrophin immunofluorescence from young (4 months old), 24-month-old untreated or 24-month-old HK-treated mice after 4 months of treatment. q, Representative pictures of H&E staining of kidney sections of young (4 months old), 24-month-old untreated and 24-month-old treated animals after 4 months of treatment. Insets show enlarged image of dashed box. r, Quantification of glomeruli diameter (Young n = 7; UT n = 6; T n = 6). s, Representative pictures of PSR staining of kidney sections of young (4 months old), 24-month-old untreated and 24-month-old treated animals after 4 months of treatment. Insets show enlarged image of dashed box. t, Quantification of percentage of fibrotic area in kidney sections (Young n = 6; UT n = 6; T n = 9). u, Kidney mRNA expression of Fn1, Col1a1 and Col3a1 (each dot represents a different animal; Young n = 8, UT n = 10, T n = 14). Data presented in d, h, i, j, k, l, n, o, r, t and u are presented as mean ± s.e.m. Data presented in e are presented as a violin plot. Statistical test used in b and c: log-rank test. Statistical test used in e, h and j: two-tailed unpaired t-test. Statistical test used in i, k, l, n, r, t and u: one-way ANOVA with Holm–Šidák’s multiple comparisons test. Statistical test used in o: Kruskal–Wallis test with Dunn’s multiple comparisons test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; NS, nonsignificant. Exact P values found in source data files.
