Fig. 5: Inhibition of ILPHF6 cells reduces stress susceptibility. | Molecular Psychiatry

Fig. 5: Inhibition of ILPHF6 cells reduces stress susceptibility.

From: Regulation of stress susceptibility by chromatin-binding protein PHF6 in the pituitary intermediate lobe

Fig. 5

a Schematic illustration of the generation of Phf6-CreER knock-in (KI) mice and the experimental strategy for labeling ILPHF6 cells through stereotactic injection of AAV-DIO-mCherry into the pituitary intermediate lobe of adult male Phf6-CreER mice, followed by tamoxifen-induced recombination. b Representative images show colocalization of endogenous PHF6 (green) and mCherry (red) in the pituitary intermediate lobe. Scale bars, 100 μm. c Quantification revealed that 97.5% ± 0.3% of mCherry-positive cells co-expressed endogenous PHF6 (n = 3). d Experimental design for ILPHF6 cell inhibition and subsequent evaluation of stress-induced anxiety-like behaviors. e Left: Representative images showing expression of hM4D-mCherry (or mCherry) in the IL of Phf6-CreER mice, co-stained with c-Fos (green). Right: Quantification demonstrated significantly reduced c-Fos expression in hM4D-expressing IL cells compared to mCherry controls after RS-stress (mCherry, n = 3; hM4D, n = 3, *** p < 0.001, unpaired Student’s t-test). DAPI is shown in blue. Scale bars, 50 μm. f Representative locomotor trajectories and quantification of time spent in the center area of the OF test. Inhibition of ILPHF6 cells significantly increased the time spent in the center area of the OF test following RS stress (mCherry-saline, n = 9; hM4D-saline, n = 10; mCherry-CNO: n = 9; hM4D-CNO: n = 10; * p < 0.05, *** p < 0.001, two-way ANOVA with Bonferroni’s multiple comparisons test). g Representative locomotor trajectories and quantification of time spent in the open arms of the EPM test. Inhibition of ILPHF6 cells significantly increased the time spent in the open arms of the EPM test after RS stress (mCherry-saline, n = 9; hM4D-saline, n = 9; mCherry-CNO: n = 9; hM4D-CNO: n = 10; * p < 0.05, ** p < 0.01, two-way ANOVA with Bonferroni’s multiple comparisons test). h Quantification of the latency to feed in the NSF test. Inhibition of ILPHF6 cells significantly decreased the latency to feed in the NSF test following RS stress (mCherry-saline, n = 9; hM4D-saline, n = 10; mCherry-CNO: n = 9; hM4D-CNO: n = 10; * p < 0.05, ** p < 0.01, two-way ANOVA with Bonferroni’s multiple comparisons test. Data are presented as mean ± SEM.

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