Fig. 2: PHF6 deficiency in ^ILPHF6 cells reduces stress susceptibility.

a Strategy for generating conditional Phf6 knockout (cKO) mice by crossing male POMC-Cre mice with female Phf6^flox/flox mice. b Representative images of endogenous PHF6 (red) and endogenous POMC (green) in the pituitary of control and Phf6 cKO mice, confirming deletion of Phf6 in cKO mice. Scale bars, 100 μm. c Experimental design for assessing anxiety-like behaviors in control and Phf6 cKO mice under basal and stress conditions. d-f Under RS-stress conditions, Phf6 cKO mice displayed increased center area time in the OF test (d), increased open-arm time in the EPM test (e), and decreased latency to feed in the NSF test (f), compared to the control mice (Control, n = 16; Phf6 cKO, n = 14; * p < 0.05, ** p < 0.01, *** p < 0.001, two-way repeated-measures analysis of variance (ANOVA) with Bonferroni’s multiple comparisons test). Example locomotor trajectories of the Phf6 cKO and control mice in the OF test (d) and EPM test (e) under RS-stress conditions are shown, with the center area (d, dotted square) and open arms (e, dotted outline) highlighted. g-i Under FS-stress conditions, Phf6 cKO mice exhibited increased center area time in the OF (g), increased open-arm time in the EPM (h), and decreased latency to feed in the NSF test (i) relative to controls (Control, n = 9; Phf6 cKO, n = 9, * p < 0.05, ** p < 0.01, two-way repeated-measures ANOVA with Bonferroni’s multiple comparisons test). Example locomotor trajectories of the Phf6 cKO and control mice in the OF test (g) and EPM test (h) under FS-stress conditions are shown. j-l Under 2MT-stress conditions, Phf6 cKO mice demonstrated increased center area time in the OF test (j), increased open-arm time in the EPM test (k), and decreased latency to feed in the NSF test (l), compared to the control mice (Control, n = 8; Phf6 cKO, n = 11, * p < 0.05, ** p < 0.01, two-way repeated-measures ANOVA with Bonferroni’s multiple comparisons test). Example locomotor trajectories of the Phf6 cKO and control mice in the OF test (j) and EPM test (k) under 2MT-stress conditions are shown. m Experimental design of chronic restraint stress (CRS) induction and assessment of depression-like behaviors in control and Phf6 cKO mice. n Quantification of the immobility duration and latency to immobility in the FST. Compared to control mice, Phf6 cKO mice exhibited decreased immobile duration and increased latency to immobility in FST under CRS conditions (n = 11 per group; * p < 0.05, two-way repeated-measures ANOVA with Bonferroni’s multiple comparisons test). o Quantification of the sucrose preference in the SPT. Compared to control mice, Phf6 cKO mice showed increased sucrose preference in SPT under CRS conditions (n = 11 per group; * p < 0.05, two-way repeated-measures ANOVA with Bonferroni’s multiple comparisons test). Data are presented as mean ± SEM.