Fig. 1 Mmp-2 activity is increased in brains after HFD intake.

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Fig. 1 Mmp-2 activity is increased in brains after HFD intake. Mmp-2 activity is increased in brains after HFD intake. (A) Representative gel zymography showing the activity of Mmp-2 in the brain of rats fed a HFD and a CD. (B) Values of the band density in (A) are presented as density units (DU). Data are shown as means ± SD (n = 8 per group). (C) Representative sections of the hypothalamus of rats fed a CD and a HFD after immunolabeling with an antibody against the active form of Mmp-2. The insets show an overview, with black squares delineating the magnified fields. Arrows point to the Mmp-2–positive cells. (D) WB analysis of Mmp-2 in brain homogenates of rats fed a HFD and a CD. Equal amounts of protein were subjected to SDS–polyacrylamide gel electrophoresis, and Mmp-2 (active form) was detected. (E) Mmp-2 and β-actin band densities were determined, and measurements were presented as relative density units (RDU) of the Mmp-2/β-actin ratio (*P < 0.05, HFD versus CD; data are shown as means ± SD; n = 9 per group). (F) Representative WB showing ObR extracellular domain signal in brains of the CD and HFD groups. (G) Optical density measurements of the WB signal and data are presented as DU (*P < 0.05, CD versus HFD; data are shown as means ± SD; n = 8). (H) IF imaging of the ObR extracellular domain in HFD and CD. White arrows point to positive staining in CD and HFD. (I) Silver stain of ObR (left), Mmp-2 (middle), and ObR incubated with Mmp-2 (right). (J) Extracellular domain of the leptin receptor. Cytokine receptor homology (CRH-2) is the main binding site for leptin. The immunoglobulin (Ig)–like and fibronectin type 3 (FN3) domains are essential for receptor signaling. Stars in the extracellular domain represent the cleavage sites determined by high-performance liquid chromatography with tandem mass spectrometry (LC-MS/MS). Rafi Mazor et al., Sci Transl Med 2018;10:eaah6324 Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works