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. Author manuscript; available in PMC: 2021 Mar 28.
Published in final edited form as: Nat Cell Biol. 2020 Sep 28;22(10):1170–1179. doi: 10.1038/s41556-020-00579-5

Figure 5. SIRT1 undergoes lysosomal degradation during aging in mouse and human.

Figure 5.

a,b, Spleens (a) and testes (b) from young (2–4 months) and aged (19–26 months) C57BL/6 mice were analyzed by western blotting and RT-qPCR. Data are mean ± s.e.m.; unpaired two-tailed Students’ t-test; n = 4 animals. c,d, Spleens (c) and testes (d) were analyzed by western blotting and RT-qPCR for SIRT1 expression, from aged (19–24 months) mice subjected to daily i.p. injection of 10 mg/kg Lys05 in PBS or PBS control in 100 μL volume for two weeks. Data are mean ± s.e.m.; two-tailed Mann-Whitney test. For spleen protein, control group n = 8 animals, Lys05 group n = 7 animals; RNA, control group n = 8 animals, Lys05 group n = 6 animals. For testis protein and RNA, control group n=6 animals, Lys05 group n=6 animals. e.HSPC populations were isolated from young (2–4 months) and aged (20–26 months) C57BL/6 mice, cultured with or without 2 μM Lys05 for 24 hours and analyzed by western blotting and RT-qPCR. For protein, data are mean ± s.e.m.; one-way ANOVA coupled with Tukey’s test; n=6 independent experiments. For RNA, data are mean ± s.e.m.; unpaired two-tailed Students’ t-test; n=4 independent experiments. f. Freshly sorted CD8+CD28+ (control) and CD8+CD28- T cells were treated with Lys05 at doses of 0 and 5 μM for 14 hours, and then were harvested and analyzed by western blotting. Donor age: 53, 54 and 66. Data are mean ± s.d.; unpaired two-tailed Students’ t-test; n = 3 human donors. In a-f, western blot quantification: SIRT1 bands were normalized to GAPDH bands, for testes both bands of SIRT1 were considered. In a-e, RT-qPCR quantification: data were normalized to 18S. Statistical information and unprocessed blots are provided as source data.