Fig. 4

ESP22-induced c-Fos expression in the amygdala-hypothalamus axis of female mice. a Representative ISH sections of the MeA from female mice stimulated with control buffer or ESP22. c-Fos cRNA probe (green) was used in conjunction with nuclear DAPI staining (blue). Abbreviations: MeApd, MeA posterodorsal part; MeApv, MeA posteroventral part; D, dorsal; and V, ventral. Scale bar, 100 µm. b Quantification of c-Fos-positive neurons in the MeA. The number of sections counted to determine the number of c-Fos-positive neurons in each brain area of each animal were: MeAa, 4; MeApd, 12; and MeApv, 12. Error bars, S.E.M. n = 4–5. MeAa, MeA anterior part. c Representative ISH sections of the BNST, as detailed in a. MA, medial division anterior part; L, lateral division; MV, medial division ventral part; LV, lateral division ventral part; MP, medial division posterior part; f, fornix; and ac, anterior commissure. Scale bar, 100 µm. Image adapted from the Allen Mouse Brain Atlas (©2004 Allen Institute for Brain Science. Allen Mouse Brain Atlas. Available from: mouse.brain-map.org). d Quantification of c-Fos-positive neurons in the BNST. Ten sections were counted for each BNST area per animal. n = 7. e Representative ISH sections of the VMH from female mice stimulated with control buffer or ESP22. SF1 cRNA probe (red) and c-Fos cRNA probe (green) were used in conjunction with nuclear DAPI staining (blue). Abbreviations: VMHd, VMH dorsal part; VMHvl, VMH ventrolateral part; D, dorsal; and L, lateral. f Quantification of c-Fos-expressing neurons in the VMH. n = 4–7; 16 sections from each animal were quantified. Error bars, S.E.M. ***p < 0.001 and *p < 0.05 by Wilcoxon rank sum test