brunello lentiviral library  (Addgene inc)

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: 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

Journal: iScience

Article Title: CCT5 maintains mitotic fidelity and promotes early colorectal tumorigenesis

doi: 10.1016/j.isci.2026.115223

Figure Lengend Snippet: CCT5 is significantly upregulated in early colorectal cancer (A) Volcano plot shows differentially expressed genes between early-stage CRC and normal tissues from GEO datasets ( GSE39582 ). (B) CCT5 was upregulated in a variety of tumors from the TCGA database. (C and D) TCGA data analysis shows that the expression of CCT5 was significantly increased in paired (left) and unpaired (right) colon cancer tissues compared with adjacent tissues. (E) Proteomic data from the CPTAC dataset (97 CRC vs. 100 normal samples) confirm elevated CCT5 protein expression in CRC. (F) Subgroup analysis shows particularly high CCT5 levels in stage I (early-stage) CRC tissues. (G) Receiver operating characteristic (ROC) curve shows that the diagnostic efficacy of CCT5 for CRC (AUC = 0.941, 95%CI: 0.919–0.963), and had high specificity and sensitivity in T1 CRC (AUC = 0.977, 95%CI: 0.945–1.000). (H and I) RT-qPCR analysis of CCT5 expression was performed in 29 paired colorectal cancer (CRC) tissues and adjacent normal tissues ( n = 29 pairs; n represents independent patient samples). Data are presented as mean ± SD. Statistical significance was determined using a paired and unpaired t test. Significance levels are indicated as ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (J and K) Western blot analysis confirms the upregulation of CCT5 in 12 paired colorectal cancer (CRC) tissues and adjacent normal tissues. Each lane represents an independent patient sample ( n = 12 pairs; n represents independent patient samples). Quantification was performed based on densitometric analysis. Data are presented as mean ± SD. Statistical significance was determined using a paired two-tailed Student’s t test. ∗ p < 0.05. (L) Quantitative real-time PCR (RT-qPCR) analysis of CCT5 mRNA expression levels in normal colon epithelial cells (NCM460) and colorectal cancer (CRC) cell lines (HCT116 and SW480). Experiments were performed in n = 3 independent biological replicates, each measured in technical triplicate. Data are presented as mean ± SD. Statistical significance was determined using an unpaired two-tailed Student’s t test. Significance levels are indicated as ∗∗ p < 0.01 and ∗∗∗∗ p < 0.0001. (M) Representative western blot showing CCT5 protein expression in normal colon epithelial cells (NCM460) and colorectal cancer (CRC) cell lines (HCT116 and SW480). GAPDH was used as a loading control. Representative immunoblots from n=3 independent biological esperiments are shown (n represents independent experiments). (N, O, P) Representative hematoxylin and eosin (H&E), immunofluorescence (IF), and immunohistochemistry (IHC) images show a progressive increase in CCT5 expression from adjacent normal tissues to early-stage colorectal cancer (EC) and advanced CRC tissues. Representative images from n = 8 independent patient samples per group are shown. Scale bars, 100 μm (N). Quantification was performed from three randomly selected fields per sample. Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test. Significance levels are indicated as ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Genetically Engineered Mouse Models (GEMMs): Heterozygous CCT5 knockout mice ( CCT5 +/– ) were purchased from Cyagen Biosciences (Cat# KOAI221117YZ1).

Techniques: Expressing, Diagnostic Assay, Quantitative RT-PCR, Western Blot, Two Tailed Test, Real-time Polymerase Chain Reaction, Control, Immunofluorescence, Immunohistochemistry

Journal: iScience

Article Title: CCT5 maintains mitotic fidelity and promotes early colorectal tumorigenesis

doi: 10.1016/j.isci.2026.115223

Figure Lengend Snippet: Loss of CCT5 suppresses epithelial proliferation and dysplastic transformation (A) Flow chart illustrates the establishment of the early-stage CRC mouse model. (B and C) Time-course analysis of body weight changes during DSS-induced tumorigenesis in the indicated genotypes. Each comparison was performed using independent mouse cohorts with their corresponding wild-type controls ( n = 7 mice per group; n represents independent animals). Data are presented as mean ± SD. Statistical analysis was performed separately for each panel using two-way repeated-measures ANOVA (genotype × time) followed by Šídák’s multiple comparisons test. DSS treatment periods are indicated. Significance levels are shown as ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (D–G) Ki-67 immunohistochemistry (IHC) analysis of colonic epithelium in wild-type (WT), heterozygous (CCT5 +/− ), and conditional knockout (CCT5 flox/+;Vil−Cre ) mice. Representative images from n = 7 independent mice per group are shown (n represents independent animals). Scale bars, 250 μm (D, E, overview) and 50 μm (D, E, magnified views). Quantification of the proliferative index was performed by calculating the percentage of Ki-67–positive epithelial cells from five randomly selected fields per mouse and averaging per mouse. Data are presented as mean ± SD. Statistical analysis was performed separately for each independent cohort and for proximal and distal regions using unpaired two-tailed t-tests. Significance levels are indicated as ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (H) Representative images showing colons from AOM/DSS-treated wild-type and CCT5-deficient mice. n=7 mice per group. (Note: Images in panel were cropped from larger microscopic fields solely for presentation clarity; no digital manipulation or rearrangement of signal content was performed). (I) Representative hematoxylin and eosin (H&E) staining of AOM/DSS-induced colon cancer tissues. Representative images from n = 7 independent mice per group are shown (n represents independent animals). Scale bars, 2.5 mm (overview) and 500 μm (magnified views). (J and K) Quantification of tumor burden in CCT5 +/− and CCT5 flox/+; Vil-Cre mice compared with their respective wild-type controls. Each data point represents the total number of tumors per individual mouse ( n = 7 independent mice per group; n represents independent animals). Data are presented as mean ± SD. Statistical analysis was performed separately for each independent cohort using unpaired two-tailed t-tests. ∗∗∗∗ p < 0.0001.

Article Snippet: Genetically Engineered Mouse Models (GEMMs): Heterozygous CCT5 knockout mice ( CCT5 +/– ) were purchased from Cyagen Biosciences (Cat# KOAI221117YZ1).

Techniques: Transformation Assay, Comparison, Immunohistochemistry, Knock-Out, Two Tailed Test, Staining

Journal: iScience

Article Title: CCT5 maintains mitotic fidelity and promotes early colorectal tumorigenesis

doi: 10.1016/j.isci.2026.115223

Figure Lengend Snippet: CCT5 deletion markedly impairs dysplastic and neoplastic progression in vivo (A) Representative hematoxylin and eosin (H&E)-stained sections show high-grade and low-grade dysplastic lesions. Representative images from n = 7 independent mice per group are shown (n represents independent animals). Scale bars, 250 μm (overview) and 50 μm (magnified views). (B–E) Quantitative analysis of lesion number and lesion area for high-grade dysplasia (HGD)/carcinoma and low-grade dysplasia (LGD) in WT and mutant mice. Each data point represents the total number (or area) of lesions per individual mouse ( n = 7 independent mice per group; n represents independent animals). Data are presented as mean ± SD. Statistical significance was determined using the unpaired two-tailed Student’s t test for comparisons between WT and mutant mice within each pathological category. ∗∗∗∗ p < 0.0001. (F and G) Immunohistochemistry (IHC) analysis of CCT5 expression in high-grade (F) and low-grade (G) dysplastic tissues compared with adjacent paratumor tissues. Representative images from n = 7 independent mice per group are shown (n represents independent animals). Scale bars, 250 μm (overview) and 50 μm (magnified views). Quantification was performed from five randomly selected fields per mouse and averaged per mouse. Data are presented as mean ± SD. Statistical significance was determined using a paired two-tailed Student’s t test. ∗ p < 0.05 and ∗∗ p < 0.01. (H and I) Immunohistochemistry (IHC) analysis of Ki67 expression in high-grade (H) and low-grade (I) dysplastic tissues compared with adjacent paratumor tissues. Representative images from n = 7 independent mice per group are shown (n represents independent animals). Scale bars, 250 μm (overview) and 50 μm (magnified views). Quantification was performed by calculating the percentage of Ki67-positive epithelial cells from multiple randomly selected fields per mouse and averaging per mouse. Data are presented as mean ± SD. Statistical significance was determined using an unpaired two-tailed Student’s t test. ∗∗∗ p < 0.001.

Article Snippet: Genetically Engineered Mouse Models (GEMMs): Heterozygous CCT5 knockout mice ( CCT5 +/– ) were purchased from Cyagen Biosciences (Cat# KOAI221117YZ1).

Techniques: In Vivo, Staining, Mutagenesis, Two Tailed Test, Immunohistochemistry, Expressing

Journal: iScience

Article Title: CCT5 maintains mitotic fidelity and promotes early colorectal tumorigenesis

doi: 10.1016/j.isci.2026.115223

Figure Lengend Snippet: Loss of CCT5 significantly inhibits the proliferation and tumorigenic capacity of CRC cells in vitro and in vivo (A and B) Validation of CCT5 knockdown at the mRNA level in HCT116 and SW480 cells by quantitative real-time PCR (RT-qPCR) and at the protein level by Western blot. Experiments were performed in n = 3 independent biological replicates, each measured in technical triplicate. Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test. ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001. (C and D) Cell viability assays show reduced proliferation upon CCT5 knockdown in HCT116 (C) and SW480 (D) cells. Experiments were performed in n = 3 independent biological replicates, each measured in technical triplicate. Data are presented as mean ± SD. Statistical significance was determined using two-way repeated-measures ANOVA followed by Šídák’s multiple comparisons test. Significance levels are indicated as ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. (E and F) Colony formation assays demonstrate reduced clonogenic growth upon CCT5 knockdown in HCT116 and SW480 cells. Experiments were performed in n = 3 independent biological replicates, each conducted in technical triplicate. Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test. Significance levels are indicated as ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. (G and H) Flow cytometry analysis of apoptosis in HCT116 (G) and SW480 (H) cells follows CCT5 depletion. Experiments were performed in n = 3 independent biological replicates (n represents independent repeats). Data are presented as mean ± SD. Statistical significance was determined using an unpaired two-tailed Student’s t test. ∗∗ p < 0.01 and ∗∗∗∗ p < 0.0001. (I) Representative images of subcutaneous xenograft tumors derived from control and CCT5-knockdown cells. Tumors from n = 10 independent mice per group are shown (n represents independent animals). (J) Representative hematoxylin and eosin (H&E)-stained sections of subcutaneous tumors derived from control and CCT5-knockdown cells. Representative sections from n = 10 independent mice per group are shown (n represents independent animals). Scale bars, 2.5 mm (overview) and 250 μm (magnified views). (K) Tumor growth curves and endpoint tumor weights show reduced tumorigenesis in CCT5-depleted groups. Tumor volume was measured at the indicated time points in n = 10 independent mice per group (n represents independent animals). Data are presented as mean ± SD. Tumor growth curves were analyzed using two-way repeated-measures ANOVA followed by Šídák’s multiple comparisons test. Endpoint tumor weights were analyzed using an unpaired two-tailed Student’s t test. ∗∗ p < 0.01 and ∗∗∗∗ p < 0.0001. (L) Western blot analysis of CCT5 expression in tumor tissues derived from control and CCT5-knockdown xenografts. Representative immunoblots from n = 3 independent mice per group are shown (n represents independent animals). Each lane represents an independent tumor sample. (M and N) Immunohistochemistry (IHC) analysis confirms reduced Ki-67 expression in tumor tissues derived from CCT5-knockdown xenografts. Representative images from n = 3 independent mice per group are shown (n represents independent animals). Scale bars, 1 mm (M, overview) and 100 μm (M, magnified views). Quantification was performed by calculating the percentage of Ki-67–positive cells from multiple randomly selected fields per tumor and averaged per mouse. Data are presented as mean ± SD. Statistical significance was determined using an unpaired two-tailed Student’s t test. ∗∗∗ p < 0.001.

Article Snippet: Genetically Engineered Mouse Models (GEMMs): Heterozygous CCT5 knockout mice ( CCT5 +/– ) were purchased from Cyagen Biosciences (Cat# KOAI221117YZ1).

Techniques: In Vitro, In Vivo, Biomarker Discovery, Knockdown, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Flow Cytometry, Two Tailed Test, Derivative Assay, Control, Staining, Expressing, Immunohistochemistry

Journal: iScience

Article Title: CCT5 maintains mitotic fidelity and promotes early colorectal tumorigenesis

doi: 10.1016/j.isci.2026.115223

Figure Lengend Snippet: Loss of CCT5 disrupts normal mitotic progression in colon cancer cells (A) Volcano plot shows differentially expressed proteins following CCT5 knockdown. Proteomic analysis was performed using label-free LC-MS/MS in n = 3 independent biological replicates per group. Differential expression analysis was conducted using two-tailed Student’s t test, and p values were adjusted using the Benjamini-Hochberg method. Proteins with |log2 fold change| > 1 and adjusted p < 0.05 were considered significantly differentially expressed. The x axis represents log2 fold change, and the y axis represents –log10 ( p value). Upregulated ( n = 202) and downregulated ( n = 168) proteins are indicated. (B) Differential proteins identified after CCT5 knockdown were most significantly enriched in cell cycle-related pathways, as revealed by KEGG analysis. (C) GSEA of differentially expressed proteins revealed significant enrichment in cell cycle and mitotic pathways. (D) Flow cytometry analysis of cell cycle distribution follows CCT5 knockdown. Experiments were performed in n = 3 independent biological replicates (n represents independent experiments). Data are presented as mean ± SD. Statistical significance for the G2/M phase fraction was determined using an unpaired two-tailed Student’s t test. ∗∗∗∗ p < 0.0001. (E and F) Immunofluorescence staining of phospho-Histone H3 (Ser10) in HCT116 cells following CCT5 knockdown. Representative images are shown (E). Scale bars, 50 μm. For each condition, three randomly selected fields were analyzed per experiment and averaged per experiment. Experiments were performed in n = 3 independent biological replicates (n represents independent experiments). Data are presented as mean ± SD. Statistical significance for the mitotic index was determined using an unpaired two-tailed Student’s t test. ∗∗ p < 0.01. (G and H) Immunofluorescence staining of β-tubulin and DAPI shows mitotic abnormalities, including chromosome misalignment and multipolar spindle formation, following CCT5 depletion in HCT116 cells (G). Representative images from n = 3 independent biological experiments are shown (n represents independent experiments). Scale bars, 2.5 μm. (H) Quantification of abnormal mitotic events based on spindle morphology and chromosome alignment. For each condition, three randomly selected fields were analyzed per experiment and averaged per experiment. Experiments were performed in n = 3 independent biological replicates. Data are presented as mean ± SD. Statistical significance for each defect type was determined using an unpaired two-tailed Student’s t test. ∗∗∗ p < 0.001. (I) Time-course immunofluorescence analysis of mitotic progression following nocodazole release in control and CCT5-depleted HCT116 cells. Cells were stained with β-tubulin to visualize spindle morphology and DAPI to assess chromatin organization. Representative images from n = 3 independent biological experiments are shown at the indicated time points (in minutes). Scale bars, 10 μm. For each experiment, mitotic cells were classified into prophase, metaphase, anaphase, or telophase based on spindle morphology and chromatin configuration. Percentages were calculated from three randomly selected fields per experiment and averaged per experiment.

Article Snippet: Genetically Engineered Mouse Models (GEMMs): Heterozygous CCT5 knockout mice ( CCT5 +/– ) were purchased from Cyagen Biosciences (Cat# KOAI221117YZ1).

Techniques: Knockdown, Liquid Chromatography with Mass Spectroscopy, Quantitative Proteomics, Two Tailed Test, Flow Cytometry, Immunofluorescence, Staining, Control

Journal: iScience

Article Title: CCT5 maintains mitotic fidelity and promotes early colorectal tumorigenesis

doi: 10.1016/j.isci.2026.115223

Figure Lengend Snippet: CCT5 depletion impairs APC/C activity and substrate degradation (A) Western blot time-course analysis of synchronized HCT116 cells reveals impaired degradation of cyclin B1 and Securin in CCT5-deficient cells. (B and C) Quantitative densitometric analysis of cyclin B1 (B) and securin (C) levels follows release from double thymidine block. Protein expression was normalized to GAPDH and expressed relative to the 0-h control condition. Experiments were performed in n = 3 independent biological replicates (n represents independent experiments). Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA (group × time) followed by Šídák’s multiple comparisons test. ∗ p < 0.05, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. (D) Immunofluorescence staining of cyclin B1 in control and CCT5-knockdown HCT116 cells. Cyclin B1 (green) and nuclei (DAPI, blue) are shown. Representative images from n = 3 independent biological experiments are presented (n represents independent experiments). Scale bars, 10 μm. The staining pattern suggests increased nuclear localization of cyclin B1 in CCT5-depleted cells. (E) CCT5 was immunoprecipitated, and the presence of CDC20, BubR1, Bub3, and Mad2L1 was assessed by immunoblotting. Representative immunoblots from n = 3 independent biological experiments are shown (n represents independent experiments). An interaction between CCT5 and CDC20 was detected, whereas no detectable interaction with BubR1, Bub3, or Mad2L1 was observed under these experimental conditions. (F) LC-MS/MS analysis of proteins co-immunoprecipitated with CCT5 identified CDC20-derived peptides. Representative MS/MS spectra of CDC20 peptides detected in CCT5 immunoprecipitates are shown. (G) Immunofluorescence staining of CCT5 (green) and CDC20 (red) in HCT116 cells. Partial overlap of CCT5 and CDC20 fluorescence signals was observed in the cytoplasm. Representative images from n = 3 independent biological experiments are shown (n represents independent experiments). Scale bars, 25 μm (overview) and 8 μm (magnified views).

Article Snippet: Genetically Engineered Mouse Models (GEMMs): Heterozygous CCT5 knockout mice ( CCT5 +/– ) were purchased from Cyagen Biosciences (Cat# KOAI221117YZ1).

Techniques: Activity Assay, Western Blot, Blocking Assay, Expressing, Control, Immunofluorescence, Staining, Knockdown, Immunoprecipitation, Liquid Chromatography with Mass Spectroscopy, Derivative Assay, Tandem Mass Spectroscopy, Fluorescence

Journal: iScience

Article Title: CCT5 maintains mitotic fidelity and promotes early colorectal tumorigenesis

doi: 10.1016/j.isci.2026.115223

Figure Lengend Snippet: CCT5 maintains the structural integrity of the MCC-APC/C complex and facilitates APC/C activation (A) Co-immunoprecipitation (CoIP) analysis of CDC20-associated proteins follows release from double thymidine block. CDC20 was immunoprecipitated, and the association of mitotic checkpoint complex (MCC) components, including BUBR1, CDC27, BUB3, and MAD2L1, was examined by immunoblotting. Representative immunoblots from n = 3 independent biological experiments are shown (n represents independent experiments). Reduced association of MCC components with CDC20 was observed in CCT5-depleted cells. (B) Co-immunoprecipitation (CoIP) analysis of APC/C-associated proteins follows release from double thymidine block. APC3 was immunoprecipitated, and the association of MCC components, including BUBR1, CDC27, CDC20, BUB3, and MAD2L1, was examined by immunoblotting. Representative immunoblots from n = 3 independent biological experiments are shown (n represents independent experiments). Reduced association of MCC components with the APC/C complex was observed in CCT5-depleted cells, while input protein levels remained comparable. (C and D) Cycloheximide (CHX) chase assay shows reduced stability of CDC20 in CCT5-deficient HCT116 cells. Representative immunoblots are shown in (C). Quantitative densitometric analysis of CDC20 protein levels normalized to GAPDH and expressed relative to the 0-h time point is shown in (D). Experiments were performed in n = 3 independent biological replicates (n represents independent experiments). Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA (group × time) followed by Šídák’s multiple comparisons test. ∗ p < 0.05, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. (E) RT-qPCR analysis of CDC20 mRNA expression follows CCT5 knockdown in HCT116 cells. No significant change in CDC20 mRNA levels was observed. Experiments were performed in n = 3 independent biological replicates (n represents independent experiments). Data are presented as mean ± SD. Statistical significance was determined using an unpaired two-tailed Student’s t test. ns, not significant. (F) Cell viability assay assesses whether CDC20 overexpression rescues the proliferation defect induced by CCT5 knockdown in HCT116 cells. Experiments were performed in n = 3 independent biological replicates (n represents independent experiments). Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA (group × time) followed by Šídák’s multiple comparisons test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. CDC20 overexpression did not restore proliferation in CCT5-depleted cells. (G and H) Flow cytometry analysis of cell cycle distribution in HCT116 cells follows CCT5 knockdown and CDC20 overexpression. Representative histograms are shown in (G). Quantification of the G2/M fraction is shown in (H). Experiments were performed in n = 3 independent biological replicates (n represents independent experiments). Data are presented as mean ± SD. Statistical significance for the G2/M fraction was determined using one-way ANOVA followed by Tukey’s multiple comparisons test. ∗∗ p < 0.01; ns, not significant. CDC20 overexpression did not restore the G2/M phase distribution in CCT5-depleted cells. (I and J) Ubiquitination assay shows increased CDC20 ubiquitination following CCT5 knockdown in HCT116 cells under asynchronous conditions (nocodazole −, MG132 +). CDC20 was immunoprecipitated, and ubiquitinated CDC20 was detected by immunoblotting with anti-ubiquitin antibodies (I). Quantitative densitometric analysis of ubiquitinated CDC20 normalized to immunoprecipitated CDC20 is shown in (J). Experiments were performed in n = 3 independent biological replicates (n represents independent experiments). Data are presented as mean ± SD. Statistical significance was assessed using an unpaired t test. ∗∗ p < 0.01. (K and L) Ubiquitination assay shows reduced CDC20 ubiquitination following CCT5 knockdown in HCT116 cells under nocodazole-induced mitotic arrest conditions (NOC +, MG132 +). CDC20 was immunoprecipitated and ubiquitinated. CDC20 was detected by immunoblotting with anti-ubiquitin antibodies (K). Quantitative densitometric analysis of ubiquitinated CDC20 normalized to immunoprecipitated CDC20 is shown in (L). Experiments were performed in n = 3 independent biological replicates (n represents independent experiments). Data are presented as mean ± SD. Statistical significance was assessed using an unpaired t test. ∗ p < 0.05.

Article Snippet: Genetically Engineered Mouse Models (GEMMs): Heterozygous CCT5 knockout mice ( CCT5 +/– ) were purchased from Cyagen Biosciences (Cat# KOAI221117YZ1).

Techniques: Activation Assay, Immunoprecipitation, Blocking Assay, Western Blot, Quantitative RT-PCR, Expressing, Knockdown, Two Tailed Test, Viability Assay, Over Expression, Flow Cytometry, Ubiquitin Proteomics

Journal: iScience

Article Title: CCT5 maintains mitotic fidelity and promotes early colorectal tumorigenesis

doi: 10.1016/j.isci.2026.115223

Figure Lengend Snippet: Schematic model: CCT5 maintains CDC20 stability and MCC-APC/C assembly to ensure mitotic progression This schematic illustrates the mechanism by which CCT5 stabilizes CDC20, regulates its ubiquitination during mitosis, facilitates its dynamic interaction with both the MCC and APC/C complexes, and ensures a timely metaphase-anaphase transition. The loss of CCT5 disrupts CDC20 stability and ubiquitination, impairs the assembly of the MCC-APC/C complex, and leads to mitotic arrest in CRC cells.

Article Snippet: Genetically Engineered Mouse Models (GEMMs): Heterozygous CCT5 knockout mice ( CCT5 +/– ) were purchased from Cyagen Biosciences (Cat# KOAI221117YZ1).

Techniques: Ubiquitin Proteomics