Fig. 2
Gli relieves aging phenotypes in vitro and in vivo. a Structure of Gli and the chemical probe of Gli (Gli-P). b MDH2 immunoblotting of proteins pulldown from in situ labeling of MRC-5 cells using Gli-P. c Binding constant (KD) of Gli to MDH2 measured by Grating-Coupled Interferometry. d SA-β-gal staining of Blk and Dox-induced senescent MRC-5 cells treated with -/Chl (200 μM)/Gli (100 μM)/Met (100 μM) for 7 days. e Quantification of d. f Relative p16INK4a level in Blk and Dox-induced senescent MRC-5 cells treated with -/Gli (100 μM)/Met (100 μM) for 7 days. g Quantification of f. h Relative MMP1 level in Blk and Dox-induced senescent MRC-5 cells treated with -/Gli (100 μM)/Met (100 μM) for 7 days. i Quantification of h. j IL-6 level in the medium of Blk and Dox-induced senescent MRC-5 cells treated with -/Gli (100 μM)/Met (100 μM) for 7 days. k IL-1β level in the medium of Blk and Dox-induced senescent MRC-5 cells treated with -/Gli (100 μM)/Met (100 μM) for 7 days. l SA-β-gal staining of MEFs (P6) treated with -/Gli (100 μM)/Met (100 μM) for 15 days. m Quantification of l. n Relative p16INK4a and γH2AX level in MEFs (P6) treated with -/Gli (100 μM)/Met (100 μM) for 5 days. o Quantification of p16INK4a in n. p Quantification of γH2AX in n. q Diagram of assays on naturally aged mice treated with -/Gli (10 mg/kg)/NMN (500 mg/kg) daily. r Lifespan curves of mice in different groups (Ctrl, n = 10; Gli, n = 12; NMN, n = 10). s Frailty index of mice in different groups (Ctrl, n = 10; Gli, n = 12; NMN, n = 10 when the test began at 18-month age). For s, every dot in the plot presents the data of 1 mouse. Error bars represent the standard deviation (± SEM.). Log-rank tests were used for analyzing the significant differences of r. Other differences were analyzed with Tukey’s multiple comparations tests (*p < 0.05, **p < 0.01, ***p < 0.005, n. s., not significant)
