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. 2020 May 11;130(6):3124–3136. doi: 10.1172/JCI135026

Figure 1. Melatonin deficiency–induced mtDNA release mediated neuroinflammation.

Figure 1

(A) LC-MS quantification of melatonin in pineal gland isolated at 4:00 a.m. from 20-week-old WT and AANAT-KO mice (n = 3). (B) Quantification of malondialdehyde (MDA), an indicator of lipid peroxidation, and (C) protein carbonylation, an indicator of protein oxidation in whole brain of 8- and 20-week-old WT and AANAT-KO mouse brain lysate (n = 3). (D) Ventricle volume in 20-week-old WT and AANAT-KO brain (n = 3). (E) qPCR of cytosolic mtDNA in 8- and 20-week-old WT and AANAT-KO brain using primers for mt-CO1, mt-Dloop1, and mt-Dloop3, mitochondrial genes. Cytosolic mtDNA is plotted relative to amount of WT brain after normalization to β-actin from the corresponding total DNA lysate, n = 3 for 8 weeks, n = 4 for 20 weeks. (F) Representative immunoblots and (G) quantitation for cGAS, STING, IRF3, caspase-1, and β-actin in WT and AANAT-KO brain lysates. β-actin was used as a loading control; data are expressed as relative to WT control (n = 4). (H) Cytokine ELISA in brain lysate of 20-week-old WT and AANAT-KO mice expressed as pg of cytokine per mg of protein lysate (n = 4). All data are expressed as mean ± SD, analyzed by Student’s t test (A, D, G, and H), or ANOVA followed by Tukey’s test (B, C, and E). *P < 0.05; **P < 0.01; ***P < 0.001.