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Figure Lengend Snippet: Melatonin‐induced change in Cal‐27 cell metabolism by upregulation of fatty acid oxidation and TCA metabolites. (A–D) Metabolomic study of intracellular levels of (A) succinic acid, (B) fumaric acid, (C) malic acid, and (D) ketoglutaric acid. (E) NADH and NAD+ levels measured using a colorimetric test and expressed as a ratio. (F–I) Western blot analysis of (F) MDAC, (G) EHHADH, and (H) PACC/ACC ratio. (J–M) Evaluation of SC formation by BNGE. Treatment groups included vehicle (control) and melatonin (aMT) at 500 or 1000 µM for 24 h (gray range) or 48 h (blue range) in Cal‐27 cells. Data are presented as mean ± standard error of the mean ( n = 3–7 for each group; one‐tailed unpaired t ‐test; * p < .05; ** p < .01; *** p < .001 vs. control). BNGE, blue native gel electrophoresis; EHHADH, enoyl‐CoA hydratase and 3‐hydroxyacyl CoA dehydrogenase; PACC/ACC, phosphorylated/no‐ phosphorylated acetyl‐CoA carboxylase; SC, supercomplex; TCA, tricarboxylic acid.

Article Snippet: NAD+ levels were measured using a oxidized and reduced nicotinamide adenine dinucleotide (NAD/NADH) assay kit (ab65348; Abcam) according to the manufacturer's instructions.

Techniques: Western Blot, One-tailed Test, Nucleic Acid Electrophoresis