Fig. 3

Increase in the number of c-Fos-positive neurons in IC of 15 days RF-mice. A, B, and C The number of c-Fos positive neurons counted within a square (500 × 500 μm) in the anterior (A), middle (B), and posterior (C) insular cortex in ad libitum feeding (AL) and 15 days of restricted feeding (RF). A significantly higher number of c-Fos was observed in RF than in AL. A–C, Left. The mean (± SEM) of c-Fos positive neurons for ten subjects are shown on the y-axis. *P < 0.05; ***P < 0.001 difference between AL mice and RF mice according to unpaired t-test with Welch’s correction. A–C, Middle. The tallied regions in AI, MI, and PI are marked with yellow squares in representative mouse brain sections counterstained with DAPI. A–C, Right. Photographs of representative sections showing c-Fos positive neurons in AI (A, Right), MI (B, Right), and PI (C, Right) from AL and RF mice. Arrows indicate representative c-Fos signals (A–C, Right). Green represents the c-Fos signal. The scale bar is 50 μm