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Thermogenesis is a promising approach to limit weight gain in response to excess nutrition. In contrast to cold-induced thermogenesis, the molecular and cellular mechanisms of diet-induced thermogenesis (DIT) have not been fully characterized. Here, we explored the response of brown adipose tissue (BAT) and beige adipose tissue to high fat diet (HFD) using proteome and phosphoproteome analysis. We observed that after HFD, DIT was only activated in BAT. Furthermore, fatty acid oxidation, tricarboxylic acid cycle, and oxidative phosphorylation were also activated in BAT. Nevertheless, most metabolic pathways downregulated in beige adipose tissue. Strikingly, we found that these metabolic changes accompanied with different variation of mitochondria between BAT and beige adipose tissue as well. HFD treatment impaired mitochondrial functions and mitochondrial protein synthesis in beige adipose tissue while it stimulated mitochondrial autophagy in BAT. Together, in BAT, HFD caused increased mitochondrial activity,