Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: Role of PVPAC-Exo-circEif3c in regulating AF biological functions and its potential mechanism. PVPAC-derived exosomal circEif3c (Exo-circEif3c) promoted AFs migration and proliferation, whereas silencing exosomal circEif3c suppresses these processes. (A) Time-course analysis of circEif3c expression in AFs after Exo-circEif3c treatment (0, 6, and 12 h; 0 h as control). (B) Stable silencing efficiency and specificity of circEif3c in AFs; Exo-siR-control served as the control. (C and D) Effects of PVPAC-Exo-siR- circEif3c-1 and -2 on AF migration and proliferation assessed by wound healing and proliferation assays. Scratch closure percentage and migrated cell numbers were quantified using ImageJ and GraphPad Prism 9.5, scale bar = 150 μm. (E) and (F) FCM analysis of AF proliferation and apoptosis following treatment with PVPAC-Exo-circEif3c, Exo-miR-96–5p, and Ad-MEOX2 interaction. (G) Western blot analysis of vimentin, PHF20L1, and MEOX2 expression in AFs under high glucose and circEif3c modulation. (H) Effects of Exo-circEif3c on the expression of vimentin, PHF20L1, MEOX2, and LC3 in AFs. GAPDH was used as a loading control. All data above are presented as mean ± SD from three independent experiments. vs. the control group, ∗P < 0.05, ∗∗P < 0.01(one-way ANOVA with Dunnett's post-hoc test), n (the number of experiments) = 3.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Derivative Assay, Migration, Expressing, Control, Western Blot

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: The regulatory mchanisms of miR-96-5p in AF biology. (A) Time-course analysis of miR-96–5p expression in AFs treated with PVPAC-Exo at 0, 6, 12, and 24 h. (B) and (C) AFs were transfected with miR-96–5p mimic and NC mimic for 24 h. Then, the migration ability of AFs through migration experiments (B) and EDU assay (C) were evaluated using Image J and GraphPad Prism 9. vs. control mimic, ∗P < 0.05, ∗∗P < 0.01, n (the number of experiments) = 3, scale bar = 150 μm. (D) Correlation analysis between extracellular ncRNA levels in culture supernatant and intracellular expression of PHF20L1 and MEOX2 by RT-qPCR. (E) Bioinformatic prediction identifying PHF20L1 and MEOX2 as potential targets of miR-96–5p. (F) Predicted miR-96–5p binding sites in the 3′UTR of PHF20L1. (G) Luciferase reporter assay displayed that miR-96–5p mimic significantly reduced luciferase activity in the PHF20L1-WT group, but not in the PHF20L1-Mut group (vs. PHF20L1-Mut, ∗∗ P < 0.01), n (the number of experiments) = 3. (H) Bioinformatics analysis indicated the miR-96–5p binding sites in the 3′UTR of MEOX2. (I) Western blot exhibited no significant change in MEOX2 protein levels upon miR-96–5p overexpression. (J) Interaction network among miR-96–5p, circEif3c, PHF20L1, and MEOX2, constructed using GEPIA, ENCORI, miRNet, NDEx, and Cytoscape. (K) Western blot analysis of PHF20L1 and MEOX2 expression in AFs transfected with control-exosome, OE-exosomes, miR-comtrol mimic, miR-96–5p mimic, and UNC1215, respectively. vs. control-exosome, miR-comtrol mimic, ∗ P < 0.05, ∗∗ P < 0.01, n (the number of experiments) = 3. (L) Predicted protein–protein interaction interface between PHF20L1 and MEOX2 using Zdock 3.0.2 and PyMOL 2.5.5. (M) Co-IP experiments confirmed an interaction between PHF20L1 and MEOX2.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Expressing, Transfection, Migration, EdU Assay, Control, Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Western Blot, Over Expression, Construct, Co-Immunoprecipitation Assay

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: CircEif3c modulates AF proliferation and migration via the miR-96-5p/PHF20L 1 /MEOX2 axis. (A–C) Cell migration and proliferation assays. AFs were transfected for 24 h with Ad-GFP, siR-circEif3c, miR-96–5p mimic, or siR-MEOX2. Migration (A) and proliferation (B) were quantified (C). (D–F) AFs were co-incubated for 48 h with control mimic, Exo-(siR-)circEif3c mimic, Exo-(siR-)miR-96–5p mimic, PVPAC-exosome (Exo-control), GW4869, or Exo-siR-pAd-MEOX2. Migration (D) and proliferation (E) were assessed (F), scale bar = 150 μm. (G) Cellular fluorescence immunolocalization. nuclei (DAPI, blue), circEif3c (Cy5, red), miR-96–5p (Cy3, orange-yellow), MEOX2 (GFP, green).Scale bar = 30 μm. The above data were presented as mean ± SD. vs. Ad-GFP group, ∗ P < 0.05, ∗∗ P < 0.01, n (the number of experiments) = 3.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Migration, Transfection, Incubation, Control, Fluorescence

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: Exosomal circEif3c/miR-96-5p/PHF20L1/MEOX2 axis drives vascular remodeling in vivo. (A) Workflow: a stable PVPAC line over-expressing circEif3c supplied exosomes (Exo-Ad-circEif3c, 10 μg/mouse) that were micro-injected into perivascular adipose tissue (PVAT) surrounding the left carotid artery for 4 weeks to initiate remodeling. Subsequently, after the model was established, treatments with (Exo)-Ad-GFP, (Exo)-Ad- circEif3c, (Exo)-Ad-miR-96–5p, and (Exo)-Ad-Meox2 were administered continuously for 2 weeks, respectively. Normal saline (NS) was used as a negative control. (B) Representative H&E-stained cross-sections and concomitant ultrasonography of the common carotid artery. Black scale bars = 50 μm, yellow scale bars = 1 mm, and white scale bars = 0.1 s. (C) Immunohistochemistry. Scale bars = 20 μm. (D) Western blotting. (E) Quantification of protein levels. (F) Tissue localization of Cy5-labeled circEif3c by immunofluorescence, scale bar = 100 μm. (G) Fluorescence intensity quantification. (H) Comparative fluorescence imaging of vascular sections: (H1) Bright-field H&E vs. dark-field GFP before and after Ad-MEOX2 transfection; Scale bars = 50 μm; (H2) DM-remodeling vs MEOX2-intervention groups. Scale bars = 30 μm. (I) Whole-animal in vivo imaging of Cy5 signal. All quantitative data above are presented as mean ± SD. vs. control, ∗ P < 0.01.∗∗ P < 0.01. n (the number of animals) = 6 in each group.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: In Vivo, Expressing, Injection, Saline, Negative Control, Staining, Immunohistochemistry, Western Blot, Labeling, Immunofluorescence, Fluorescence, Imaging, Transfection, In Vivo Imaging, Control

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: Schematic illustration of the PVPAC-Exo mediated circEif3c/miR-96–5p/PHF20L1/MEOX2 axis regulating vascular remodeling.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques:

Journal: Molecular Therapy Advances

Article Title: Using the nose as a factory to secrete proteins into the lungs or circulation

doi: 10.1016/j.omta.2026.201733

Figure Lengend Snippet: GM-CSF transgene and aPAP mouse phenotype lung-level quantification at 2.5 weeks (100 μL and 10 × 5 μL treated mice given 2.3e8TU vector) and 2 months (5 × 5 μL treated mice given 6e7TU vector) post intranasal rSIV.F/HN treatment (A) Bronchoalveolar lavage fluid (BALF) GM-CSF levels measured by ELISA (Kruskal-Wallis, n = 6–15, p < 0.0001: Dunn’s post-hoc test) (data are represented as individual values with median; dotted line = lowest experimental standard of 7.8 pg/mL). (B) BALF turbidity measured as optical density (OD: 600 nM) (ANOVA, n = 6–15, p < 0.0005: Tukey’s post-hoc test) (data are represented as individual values with median). (C) Pulmonary alveolar surfactant (PAS) staining example images from no-dose and 100 μL bolus-treated mouse lungs (left) and image analysis quantification of all left-lung lobes (right) (Welch’s ANOVA, n = 6–15, p < 0.005: Dunnett’s post-hoc test) (data are represented as individual values with mean ± SD). (D) Example images showing how lung consolidation was measured from PAS-stained whole left-lung coronal sections (left), using image analysis to measure whole-lung area (middle) and consolidation area (right). (E) Consolidation area, as a percentage of total tissue area, plotted across all left-lung samples (Kruskal-Wallis, n = 6–15, p < 0.0001: Dunn’s post-hoc test) (data are represented as individual values with median). ∗ = p < 0.05; ∗∗ = p < 0.005; ∗∗∗ = p < 0.0005; ∗∗∗∗ = p < 0.0001.

Article Snippet: In this study arm, male and female GM-CSF knockout mice (B6.129S-Csf2 tm1Mlg /J, Jackson Laboratory, Bar Harbor, Maine, USA) of approximately 6 months of age were intranasally administered viral vector diluted in TSSM to achieve target total doses.

Techniques: Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Staining

Journal: Neurobiology of Stress

Article Title: USP11 drives stress-induced synaptic structural deficits and depression-like behaviors through GSK3β/mTOR signaling

doi: 10.1016/j.ynstr.2026.100791

Figure Lengend Snippet: Chronic unpredictable mild stress promotes depression-like behaviors and upregulates USP11 in mouse prefrontal cortex. (A) Schematic overview of the experimental timeline: male C57BL/6J mice underwent 1-week adaptation, followed by 4 weeks of chronic unpredictable mild stress (CUMS) and subsequent behavioral tests. (B-E) SPT, OFT, FST, TST results in control (Ctrl) and CUMS groups (n = 8, SPT, Welch's t -test, p = 0.0204; OFT, p = 0.0101; FST, p = 0.0020; TST, p = 0.0078). (F) Western blot of p-mTOR (Ser2448) (289 kDa), total mTOR (289 kDa), p-GSK3β(Ser9) (47 kDa), total GSK3β (47 kDa), and Tubulin (55 kDa) in mPFC tissue (n = 6). (G) Quantification of p-mTOR/t-mTOR and p-GSK-3β/t-GSK-3β ratios (p-mTOR, Welch's t -test, Pp= 0.0023; p-GSK-3β, p = 0.0075). (H) Western blot of USP11 (110 kDa) and Tubulin (55 kDa) in mPFC (n = 6). (I) Quantification of USP11 protein normalized to Tubulin (p = 0.002). (J) Representative immunofluorescence images for DAPI (blue, nuclear stain), USP11 (red), and merged panels in mPFC of control and CUMS mice. Scale bar: 50 μm. (K) Mean USP11 immunofluorescence intensity quantification (n = 3, p = 0.0142). Data are shown as mean ± SEM. Statistical analysis used two-tailed unpaired Student's t-test unless otherwise indicated. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The USP11 knockout (USP11-KO) mice in C57BL/6N background were generated by Cyagen Company (Suzhou, China).

Techniques: Control, Western Blot, Immunofluorescence, Staining, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: USP11 drives stress-induced synaptic structural deficits and depression-like behaviors through GSK3β/mTOR signaling

doi: 10.1016/j.ynstr.2026.100791

Figure Lengend Snippet: USP11 directly interacts with GSK3β. (A) Volcano plot of proteins detected after USP11 immunoprecipitation from mouse mPFC. Log2 fold change (x-axis) shows enrichment versus control; log2 intensity (y-axis) reflects normalized quantitation in experimental samples. USP11 served as bait; GSK3β is highlighted as an interactor (log2 intensity USP11 = 22.9, log2FC = 2.38). (B) Immunoprecipitation (IP) with anti-USP11 antibody, immunoblot (IB) detection for USP11 (110 kDa) and GSK3β (47 kDa). IP with anti-GSK3β or anti-USP11 antibody. Input: whole lysate; IgG: isotype control. (C) Validation in HEK293T transfection system: lysates of vector control or Flag-USP11 transfected cells (Flag tag, 110 kDa) subjected to IP (anti-GSK3β), IB for anti-USP11. (D) Cell lysate analysis of HEK293T single His-GSK3β, single Flag-USP11, or co-transfected groups, immunoblotted for His-GSK3β (47 kDa) and Flag-USP11 (110 kDa). (E, F) Reciprocal Co-IP verification from HEK293T co-transfection. Immunoblot analysis for His and Flag tag in His-GSK3β, Flag-USP11, and co-transfected samples. (E) Lane 1: His-GSK3β group (IP-His), Lane 2: Flag-USP11 group (IP-Flag), Lane 3: Co-transfection (IP- His) (F) Lane 1: His -GSK3β group (IP- His), Lane 2: Flag-USP11 group (IP-Flag), Lane 3: Co-transfection (IP-Flag). (G) Dot blot analysis showing specific binding between USP11 and GSK3β. BSA (100/200/500 ng) served as negative control, and purified USP11 (100/200/500 ng) was spotted on the same nitrocellulose membrane. After incubation with GSK3β protein solution, binding was detected by fluorescence imaging. (H) Immunofluorescence analysis of co-localization: Exogenous expression in HEK293T cells demonstrates USP11 (red) and GSK3β (green); endogenous expression verified in primary neurons. Nuclei stained with DAPI (blue), scale bar = 25 μm. (I) Fluorescence intensity profiles along linear ROIs: Gray values of USP11 (red) and GSK3β (green) measured with ImageJ. Dual-channel curves plotted in GraphPad Prism using exported data. (J) Pearson's correlation scatter plots for USP11(red) and GSK3β(green) fluorescence, generated using ScatterJ plugin for ImageJ. Pearson's r value shown. (K) Schematic of Flag-tagged USP11 fragment constructs used for pulldown mapping. (L) HEK293T cells were co-transfected with Flag-USP11 or its deletion mutant and His- GSK3β, followed by immunoprecipitation and immunoblot analysis for Flag and His. (M) Computational molecular docking predicts multiple direct contact sites between USP11 and GSK3β.

Article Snippet: The USP11 knockout (USP11-KO) mice in C57BL/6N background were generated by Cyagen Company (Suzhou, China).

Techniques: Immunoprecipitation, Control, Quantitation Assay, Western Blot, Biomarker Discovery, Transfection, Plasmid Preparation, FLAG-tag, Co-Immunoprecipitation Assay, Cotransfection, Dot Blot, Binding Assay, Negative Control, Purification, Membrane, Incubation, Fluorescence, Imaging, Immunofluorescence, Expressing, Staining, Generated, Construct, Mutagenesis

Journal: Neurobiology of Stress

Article Title: USP11 drives stress-induced synaptic structural deficits and depression-like behaviors through GSK3β/mTOR signaling

doi: 10.1016/j.ynstr.2026.100791

Figure Lengend Snippet: USP11 regulates GSK3β ubiquitination, phosphorylation, and synaptic protein homeostasis in neural cells (A) Western blot analysis of GSK3β ubiquitination in HEK293T cells co-transfected with Flag-vector (control), Flag-USP11 (wild-type, 110 kDa), or Flag-USP11-C318S (catalytically inactive mutant). Endogenous GSK3β and phosphorylated GSK3β at Ser9 were immunoprecipitated from cell lysates using anti-GSK3β antibody, and ubiquitination levels were detected by immunoblotting with anti-ubiquitin antibody. GSK3β: 47 kDa; ubiquitin bands detected as smear. (B) Western blot analysis of GSK3β phosphorylation in three 293T cell groups: wild-type (Ctrl), stable USP11-overexpressing line generated by lentiviral transduction (USP11-OE), and USP11-overexpressing cells subjected to siRNA knockdown (USP11-OE + siUSP11). siUSP11 was transfected to silence USP11 in the stable overexpressing cell line. Whole cell lysates were analyzed for endogenous USP11 (110 kDa), phosphorylated GSK3β at Ser9 (p-GSK3β, 47 kDa), total GSK3β (47 kDa), and GAPDH (35 kDa) as loading control. Representative results from n = 3 biological replicates per group. (C) Gray value quantification of p-GSK3β/t-GSK3β in 293T cells (n = 3, F (2, 6) = 35.38, p = 0.0005). (D) Western blot analysis of USP11 (110 kDa), phosphorylated mTOR (p-mTOR, Ser2448, 289 kDa), total mTOR (289 kDa), p-GSK3β (Ser9, 47 kDa), total GSK3β (47 kDa), and Tubulin (55 kDa) in primary neurons upon USP11 siRNA knockdown (n = 3). (E, F) Gray value quantification of p-GSK3β/t-GSK3β, and p-mTOR/t-mTOR ratios in neurons upon USP11 siRNA knockdown (n = 3, p-GSK3β, p = 0.0213, p-mTOR, p = 0.0047). (G) Immunoblot of USP11 (110 kDa), p-GSK3β (Ser9, 47 kDa), total GSK3β (47 kDa), SYN (77 kDa), and Tubulin (55 kDa) in primary neurons infected with adeno-associated virus (AAV) (n = 3). (H, I) Gray value quantification of p-GSK3β/t-GSK3β, and SYN/Tubulin ratios in neurons transduced with vector or AAV-USP11 viruses (n = 3, p-GSK3β, p = 0.0078, SYN, Welch's t -test, p = 0.0031). (J) Representative immunofluorescence of primary neurons transduced with vector or AAV-USP11 viruses, showing DAPI (blue, nuclei), SYN (green, synaptophysin), and USP11 (magenta); merged panels display synapse integrity. Scale bar: 50 μm. (K, L) Quantitative analysis from three independent biological replicates in primary neurons transduced with vector or AAV-USP11 viruses (K) Mean USP11 immunofluorescence intensity (p = 0.0416), (L) Mean SYN immunofluorescence intensity (p = 0.0035). Data are shown as mean ± SEM. Determined by t -test (baseline comparisons) or one-way ANOVA (multiple groups) unless otherwise indicated. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The USP11 knockout (USP11-KO) mice in C57BL/6N background were generated by Cyagen Company (Suzhou, China).

Techniques: Ubiquitin Proteomics, Phospho-proteomics, Western Blot, Transfection, Plasmid Preparation, Control, Mutagenesis, Immunoprecipitation, Generated, Transduction, Knockdown, Infection, Virus, Immunofluorescence

Journal: Neurobiology of Stress

Article Title: USP11 drives stress-induced synaptic structural deficits and depression-like behaviors through GSK3β/mTOR signaling

doi: 10.1016/j.ynstr.2026.100791

Figure Lengend Snippet: USP11 knockout alleviates stress-induced depressive-like behaviors and associated with mTOR Signaling (A) Western blot analysis of USP11 (110 kDa), p-mTOR (Ser2448, 289 kDa), total mTOR (289 kDa), p-GSK3β (Ser9, 47 kDa), total GSK3β (47 kDa), PSD95 (95 kDa), and Tubulin (55 kDa) in mouse mPFC from wild-type (WT) and USP11 knockout (USP11 −/− ) male mice (n = 6, Tubulin as loading control). (B–E) Quantification of baseline protein band intensity in wild-type control (WT-CON) and USP11 knockout control (KO-CON) groups: (B) USP11 (relative to Tubulin, p < 0.0001), (C) p-GSK3β (relative to total GSK3β, p = 0.0072), (D) p-mTOR (relative to total mTOR, p = 0.0028), (E) PSD95 (relative to Tubulin, p = 0.0159). n = 6/group. (F–I) Behavioral results for four groups: WT-CON, KO-CON, WT-CUMS, and KO-CUMS (OFT, distance [cm], F [3, 28] = 8.234, p = 0.0004; OFT, velocity [cm/s], F [3, 28] = 8.233, p = 0.0004; FST, F [3, 28] = 8.721, p = 0.0003; TST, F [3, 29] = 5.378, p = 0.0046). n = 8/group. (J) Western blot analysis of USP11 (110 kDa), p-mTOR (Ser2448, 289 kDa), total mTOR (289 kDa), SYN (synaptophysin, 77 kDa), and Tubulin (55 kDa) in mPFC from all four groups (n = 3). (K-M) Quantification of (K) USP11 (relative to Tubulin, F (3, 8) = 139.5, p < 0.0001), (L) p-mTOR (relative to total mTOR, F (3, 8) = 8.298, p = 0.0077), (M) SYN (relative to Tubulin, F (3, 8) = 8.811, p = 0.0065). n = 3/group. (N) Schematic overview of the experimental design, including a 7-day acclimation period, a 28-day chronic unpredictable mild stress (CUMS) procedure, the rapamycin dosing regimen (3 mg/kg, i.p., three times per week; from day 14 of CUMS until 24 h before tissue collection), and the behavioral test battery in male USP11 −/− mice. (O-R) Behavioral results for three groups in USP11 −/− mice: CON + Veh, CUMS + Veh and CUMS + Rapa. (SPT, F (2, 18) = 7.019, p = 0.0056; OFT, center time [s], F [2, 18] = 8.788, p = 0.0022; OFT, velocity [cm/s], F [2, 18] = 0.09090, p = 0.9135; TST, F [2, 18] = 7.797, p = 0.0036). n = 7/group.) (T) Quantification of p-mTOR (relative to total mTOR, F (2, 6) = 38.49, p = 0.0004) Data are shown as mean ± SEM. Determined by t -test (baseline comparisons) or one-way ANOVA (multiple groups) unless otherwise indicated. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (S) Representative immunoblots of p-mTOR (Ser2448, 289 kDa), total mTOR (289 kDa) in USP11 −/− mice under the indicated conditions. (n = 3, Tubulin as loading control).

Article Snippet: The USP11 knockout (USP11-KO) mice in C57BL/6N background were generated by Cyagen Company (Suzhou, China).

Techniques: Knock-Out, Western Blot, Control, Battery

Journal: Neurobiology of Stress

Article Title: USP11 drives stress-induced synaptic structural deficits and depression-like behaviors through GSK3β/mTOR signaling

doi: 10.1016/j.ynstr.2026.100791

Figure Lengend Snippet: Ultrastructural and dendritic morphological analysis reveals preservation of synaptic integrity and neuronal complexity in USP11 knockout mice under chronic stress (A) Representative transmission electron micrographs of the prefrontal cortex from WT-CON, KO-CON, WT-CUMS, and KO-CUMS mice, showing typical synaptic structures. Scale bar: 2 μm. (B) Quantification of synapse number per field from electron micrographs (n = 3, F (3, 8) = 33.5, p < 0.0001). Synaptic density was significantly reduced in WT-CUMS compared to WT-CON, while KO-CUMS mice showed partial rescue. (C) Measurement of postsynaptic density (PSD) thickness (nm) using ImageJ Pro Plus software on high-resolution electron micrographs (n = 3, 3 synapses per mouse, Brown-Forsythe ANOVA test, p = 0.0003). (D) Golgi staining images of prefrontal cortical neurons (magnifications: 20 × , 60 × , 100 × ) displaying dendritic arborization and spine morphology for each group. Scale bar: 50 μm. Representative circular diagrams illustrate dendritic arborization complexity of typical prefrontal cortical neurons in each group (spacing: 5 μm) for visualization. (E) Quantification of dendritic spine density (spines/μm) from Golgi-stained neurons (n = 3, F (3, 8) = 13.33, p = 0.0018). Each data point represents one independent biological sample, calculated as the within-mouse mean of spine measurements from 5 randomly selected neurons. (F, G) Sholl analysis of Golgi-stained mPFC neurons. (F) The number of dendritic intersections as a function of radial distance from the soma (step size = 10 μm; maximum radius = 120 μm). Statistical analysis was performed using two-way repeated-measures ANOVA (group × radius), followed by post hoc multiple comparisons, as appropriate. (Distance: F (1.590, 13.88) = 45.80, p < 0.0001; Treatment: F (1.257, 40.21) = 39.54, p < 0.0001, Asterisks denote P values for WT-CUMS vs. KO-CUMS at each radius.) (G) Area under the Sholl curve was calculated from the intersection–radius profiles as an integrated, single-metric summary of overall dendritic arbor complexity. (n = 3, F (3, 8) = 40.72 p < 0.001, each data point represents one independent biological sample, calculated from 5 randomly selected neurons.) Data are shown as mean ± SEM, statistical analysis by one-way ANOVA with Tukey's post hoc test unless otherwise indicated. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The USP11 knockout (USP11-KO) mice in C57BL/6N background were generated by Cyagen Company (Suzhou, China).

Techniques: Preserving, Knock-Out, Transmission Assay, Software, Staining

Journal: Neurobiology of Stress

Article Title: USP11 drives stress-induced synaptic structural deficits and depression-like behaviors through GSK3β/mTOR signaling

doi: 10.1016/j.ynstr.2026.100791

Figure Lengend Snippet: USP11 knockout conferred synaptic protection under chronic stress. (with Figdraw).

Article Snippet: The USP11 knockout (USP11-KO) mice in C57BL/6N background were generated by Cyagen Company (Suzhou, China).

Techniques: Knock-Out