Fig. 4

Increases in the number of c-Fos-positive and c-Fos-orexin-double-positive neurons in LH of 15 days RF-mice. A and B The number of c-Fos positive neurons (A) and the ratio of c-Fos-orexin double-positive neurons to the orexin neurons (B) counted within a rectangle (1200 × 400 μm) in the lateral hypothalamus (LH) in ad libitum feeding (AL) and 15 days of restricted feeding (RF). RF group showed significantly higher values. A and B, Left. The mean (± SEM) of c-Fos positive neurons (A, Left) and c-Fos-orexin double-positive neurons ratio in orexin neurons (B, Left) for ten subjects are shown on the y-axis. ****P < 0.0001 difference between AL mice and RF mice according to unpaired t-test with Welch’s correction. A and B, Middle. The tallied region in LH is marked with a yellow rectangle in the representative section of the mouse brain counterstained with DAPI. A and B, Right. Photographs of representative sections showing c-Fos positive neurons in LH (A, Right) and c-Fos positive orexin neurons in LH (B, Right) from AL and RF mice. Arrows indicate representative c-Fos signals (A, Right) and representative c-Fos-orexin signals (B Right). Green represents the c-Fos signal, and red represents the orexin signal. The scale bar is 50 μm