Fig. 1: PHF6 marks the pituitary IL cells that respond to stress.

a Representative images of endogenous PHF6 (red) and DAPI (blue) in the pituitary of male and female wild type (WT) mice at postnatal day 7 (P7), P14, P28, and P60. AL, anterior lobe; IL, intermediate lobe; PL, posterior lobe. Scale bars, 100 μm. b Schematic diagram of the pituitary intermediate lobe (IL). c Quantification of PHF6⁺/DAPI⁺ cells in the IL of adult male and female mice (male, n = 9; female, n = 5). d Representative images showing colocalization of endogenous PHF6 (red) and endogenous POMC (green) in the IL of adult WT male mice. Scale bars, 100 μm (low magnification) and 20 μm (high magnification). e Quantification of PHF6⁺/POMC⁺ cells in the IL of adult male and female mice (male, n = 9; female, n = 5). f, i, l Experimental design for c-Fos staining following a 10-min stress exposure using one of the three paradigms: restraint stress (RS) (f), forced swim (FS) (i), or predator odor 2-methyl-2-thiazoline (2MT) (l). g, j, m Representative images of c-Fos (green) and DAPI (blue) staining in the pituitary under RS-stress (g), FS-stress (j), and 2MT-stress (m) conditions, along with their respective basal controls. Scale bars, 100 μm. h Quantification of the number of c-Fos-positive cells in the IL under basal and RS-stress conditions (Base: n = 4; RS-Stress: n = 5, **** p < 0.0001, unpaired Student’s t-test). k Quantification of the number of c-Fos-positive cells in the IL under basal and FS-stress conditions (Base: n = 3; FS-Stress: n = 4, *** p < 0.001, unpaired Student’s t-test). n Quantification of the number of c-Fos-positive cells in the IL under basal and 2MT-stress conditions (Base: n = 3; 2MT-Stress: n = 5, *** p < 0.001, unpaired Student’s t-test). Data are presented as mean ± SEM.