Journal: Science Advances
Article Title: Citrulline regulates macrophage metabolism and inflammation to counter aging in mice
doi: 10.1126/sciadv.ads4957
Figure Lengend Snippet: ( A ) Graphic illustration of the workflow to profile age-associated metabolites in brain tissue, liver tissue and serum from mice at different ages using LC-MS. Age-associated metabolites were screened by Kruskal-Wallis test ( P < 0.05) and Spearman correlation ( P < 0.05). w, weeks; m / z , mass/charge ratio. ( B ) Pathway enrichment analyses of age-associated metabolites from mouse brain, liver, and serum (hypergeometric test, *** P < 0.001, ** P < 0.01, and * P < 0.05). TCA, tricarboxylic acid. ( C ) Age-associated metabolites were screened by Kruskal-Wallis test ( P < 0.05) and Spearman correlation ( P < 0.05). Red dots (brain, n = 26; liver, n = 46; serum, n = 21) and blue dots (brain, n = 49; liver, n = 60; serum, n = 59) represent metabolites that were increased and decreased over the age, respectively. Black dots represent unchanged metabolites. ( D ) Venn diagram for the overlap of age-associated metabolites in mouse brain, liver, and serum obtained from (C). ( E ) The spearman factors of seven common age-associated metabolites in mouse brain, liver, and serum datasets. Red and blue bars represent metabolites that were increased and decreased over the age, respectively. n = 3. Three dots for each metabolite represent their Spearman factors with age obtained from mouse brain, liver, and serum datasets, respectively. ( F ) Relative abundances of citrulline in mouse brain, liver, and serum at different ages. Bars represent means ± SEM. n = 8 to 10; all numbers are biologically independent samples. *** P < 0.001, ** P < 0.01, and * P < 0.05; n.s., not significant (two-tailed Student’s t test).
Article Snippet: The following primary antibodies were used: mouse β-actin (Proteintech, #66009-1-Ig), rabbit p-S6(Ser 240/244 ) [Cell Signaling Technology (CST), #5364S], rabbit HIF-1α (CST, #36169T), rabbit p16 (Abcam, #ab211542), rabbit p21 (Abcam, ab188224), mouse Iba1 (Abcam, #ab283342), mouse γH2AX (CST, #80312), rabbit p-mTOR(Ser 2448 ) (Proteintech, #67778-1-lg), mouse mTOR (Proteintech, #66888-1-lg), rabbit 4EBP1 (Proteintech, #60246-1-lg), rabbit p-4EBP1(Thr 37/46 ) (CST, #2855T), rabbit p70(S6K) (Proteintech, #14485-1-AP), rabbit p-p70(S6K)(Thr 389 ) (CST, #9234T), rabbit TNFα (Proteintech, #17590-1-AP), and rabbit IL-1β (CST, 12242S).
Techniques: Liquid Chromatography with Mass Spectroscopy, Two Tailed Test