Localization of PLAGL1 in murine hypocretin neurons under several conditions. (A-G) Murine HCRT was visualized by staining with Alexa Fluor® 488 (green) and murine PLAGL1 was visualized by staining with Alexa Fluor® 594 (red). (A) Murine hypothalamic sections at ZT6. The lateral hypothalamic area under (B) ZT6, (C) ZT18 and (D) SD6h conditions, with a relatively specific and uneven distribution of PLAGL1 (arrows). The dorsomedial hypothalamic nucleus under (E) ZT6, (F) ZT18 and (G) SD6h conditions. (H) DAPI-labeled nuclei at ZT6. (I) Alexa Fluor®488-labeled murine HCRT at ZT6. (J) Alexa Fluor® 594-labeled murine PLAGL1 at ZT6. (K) Merged image of (H, blue), (I, green) and (J, red) immunofluorescence at ZT6. (L) DAPI-labeled nuclei at SD6h. (M) Alexa Fluor®488-labeled murine HCRT at SD6h. (N) Alexa Fluor®594-labeled murine PLAGL1 at SD6h. (O) Merged image of (L, blue), (M, green), and (N, red) immunofluorescence at SD6h. Scale bar, (A) 500 µm, (B-G) 40 µm, (O) 10 µm. (H-O) These images are displayed at the same magnification. 3vt, third ventricle; Amy, amygdala; ARC, arcuate nucleus; DMH, dorsomedial hypothalamic nucleus; HCRT, hypocretin; LH, lateral hypothalamus; PLAGL1, pleomorphic adenoma gene-like 1; SD6h, 6 h of sleep deprivation; VMH, ventromedial nucleus of the hypothalamus; ZT, Zeitgeber time.