Western blot and immunohistochemistry of muscarinic acetylcholine receptor subtype 5 (mAChR5). Samples of the frontal cortex (A) and hippocampus (C) from rats fed with a standard diet (CHOW rats) and high-fat diet (DIO rats) for 5 and 17 weeks were immunoblotted using specific anti- mAChR5. Graphs report the densitometric data with CHOW rats as control, and β-actin was used as a reference loading protein. Data are mean ± S.E.M. CHOW rats 5 weeks n = 8; DIO rats 5 weeks n = 8; CHOW rats 17 weeks n = 8; DIO rats 17 weeks n = 9. Representative pictures of CHOW and DIO rats frontal cortex after 5 weeks (B) and hippocampus, CA3 subfield, after 17 weeks (D) of diet. V, fifth layer of frontal cortex; P, pyramidal neurons; R, stratum radiatum. Scale bar: 25 µm.