Journal: Cell discovery

Article Title: Psychological stress-induced systemic corticosterone directly sabotages intestinal stem cells and exacerbates colitis.

doi: 10.1038/s41421-025-00796-y

Figure Lengend Snippet: Fig. 5 FKBP5 is upregulated by corticosterone in ISCs. a Schematic diagram of RNA-seq analysis of ISCs isolated from the small intestine of Ctrl, restraint stress-treated, vehicle-treated and CORT-treated mice. Lgr5-EGFP-IRES-creERT2 mice were exposed to restraint stress or CORT treatment for 1 day (n = 3). The small intestinal crypt fractions were isolated and dissociated into single-cell suspensions. ISCs were sorted by using flow cytometry and subjected to RNA-seq analysis. b Heatmap showing the expression of stem cell marker genes, Wnt-related genes and proliferation-related genes in ISCs isolated from the small intestine of Ctrl, restraint stress-treated, vehicle-treated and CORT-treated mice. c Gene set enrichment analysis (GSEA) of transcriptome profiles showing high enrichment of FoxO signaling pathway-related genes in ISCs from the stressed (vs. Ctrl) and CORT-treated (vs. vehicle) mice. Differentially expressed gene, > 2-fold; Padj value < 0.05. d Venn diagram showing overlapping upregulated genes in ISCs from the stress-treated (vs. Ctrl) and CORT-treated (vs. vehicle) mice. e Top 5 upregulated genes in ISCs from the stress-treated (vs. Ctrl) and CORT-treated (vs. vehicle) mice. f Volcano plots showing the genes differentially expressed in ISCs from the stress-treated (vs. Ctrl) and CORT-treated (vs. vehicle) mice. g RT–qPCR analysis of Fkbp5 expression in the small intestinal crypts from the 1-day restraint stress- and CORT-treated mice. n = 6 and 9 mice for the restraint stressed and CORT-treated groups, respectively. h Western blotting for FKBP5 in the small intestinal crypts from the 2-day stress- and CORT-treated mice. β-actin was used as a loading control. i RT–qPCR analysis of Fkbp5 expression in intestinal organoids after ex vivo treatment with 200 μM CORT or 200 μM CORT simultaneously supplemented with 5 μM RU486 (n = 3). j A chromatin immunoprecipitation assay was performed using an antibody against NR3C1 on crypts isolated from the WT mice treated with vehicle or CORT for 1 day (n = 4). IgG was used as a negative control, and the enrichment of NR3C1 binding to the Fkbp5 promoter was quantified using qPCR. The data are presented as the mean ± SD. The statistical analysis was performed by unpaired two-tailed Student’s t-test for normally distributed data or the Mann‒Whitney test for non-normally distributed data and by one-way ANOVA with Dunnett’s multiple comparisons test or the Kruskal‒Wallis test with Dunn’s multiple comparisons test for non-normally distributed data. ns, p ≥0.05, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The primary antibodies used in this study included rabbit monoclonal anti-GR (NR3C1) (Cell Signaling Technology, 12041S), mouse monoclonal anti-FKBP5 (Cell Signaling Technology, 12210S), rabbit polyclonal anti-Akt (Cell Signaling Technology, 9272S), rabbit monoclonal anti-phospho-Akt (Ser473) (Cell Signaling Technology, 4060S), rabbit polyclonal anti-FOXO1 (Proteintech, 18592-1- AP), mouse monoclonal anti-beta actin (Proteintech, HRP-66009), mouse monoclonal anti-alpha tubulin (Proteintech, HRP-66031) and rabbit polyclonal anti-lamin B1(Proteintech, 12987-1-AP).

Techniques: RNA Sequencing, Isolation, Cytometry, Expressing, Marker, Quantitative RT-PCR, Western Blot, Control, Ex Vivo, Chromatin Immunoprecipitation, Negative Control, Binding Assay, Two Tailed Test

Journal: Cell discovery

Article Title: Psychological stress-induced systemic corticosterone directly sabotages intestinal stem cells and exacerbates colitis.

doi: 10.1038/s41421-025-00796-y

Figure Lengend Snippet: Fig. 7 Nr3c1 deficiency in ISC mitigates the stress-exacerbated colitis. a Schematic of the dextran sulfate sodium (DSS)-induced colitis model. WT and Nr3c1 cKO mice received a 5-day tamoxifen (TAM, 2 mg/25 g body weight) injection. A subset of the mice was were subjected to CRS treatment. During the last week of the three-week CRS treatment, mice were given 2.0% DSS in drinking water for seven days, followed by a 2-day recovery period with normal drinking water. b, c Body weight (b) and disease activity index (DAI) (c) of WT (Nr3c1fl/fl) + Ctrl (n = 8), WT + CRS (n = 9), Nr3c1 cKO (Lgr5;Nr3c1fl/fl) + Ctrl (n = 8) and Nr3c1 cKO + CRS (n = 9) mice during 2.0% DSS treatment. Body weight was monitored daily and expressed as a percentage of the initial body weight. d, e Representative image of colon (d) and quantification of colon length (e) of WT + Ctrl, WT + CRS, Nr3c1 cKO + Ctrl, Nr3c1 cKO + CRS mice (n = 8) after 7-day DSS treatment and 2-day recovery period. f‒h Representative H&E staining image (f), epithelial damage score (g), and inflammatory infiltrate score (h) of colonic section from WT + Ctrl (n = 8), WT + CRS (n = 9), Nr3c1 cKO + Ctrl (n = 7), Nr3c1 cKO + CRS (n = 9) mice after 7 days of DSS treatment and a 2-day recovery period. Scale bar: 50 µm. i Representative image of in situ hybridization for Lgr5 combined with immunofluorescence staining for Ki67 and the epithelial marker cadherin (E-cadherin) in the colons of Ctrl and CRS-treated mice with 2.0% DSS treatment. Scale bar: 50 µm. j, k Quantification of percentage of Lgr5+ cells per crypt (j), and the proportion of Lgr5+ cells expressing Ki-67 (k) in the colons of WT + Ctrl (n = 5), WT + CRS (n = 6), Nr3c1 cKO + Ctrl (n = 5) and Nr3c1 cKO + CRS (n = 6). The data are presented as the mean ± SD. The statistical analysis was performed by an unpaired two-tailed Student’s t-test for normally distributed data or the Mann‒Whitney test for non-normally distributed data. ns, p ≥0.05, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The primary antibodies used in this study included rabbit monoclonal anti-GR (NR3C1) (Cell Signaling Technology, 12041S), mouse monoclonal anti-FKBP5 (Cell Signaling Technology, 12210S), rabbit polyclonal anti-Akt (Cell Signaling Technology, 9272S), rabbit monoclonal anti-phospho-Akt (Ser473) (Cell Signaling Technology, 4060S), rabbit polyclonal anti-FOXO1 (Proteintech, 18592-1- AP), mouse monoclonal anti-beta actin (Proteintech, HRP-66009), mouse monoclonal anti-alpha tubulin (Proteintech, HRP-66031) and rabbit polyclonal anti-lamin B1(Proteintech, 12987-1-AP).

Techniques: Injection, Activity Assay, Staining, In Situ Hybridization, Marker, Expressing, Two Tailed Test