Journal: Clinical Science (London, England : 1979)

Article Title: Unravelling the effects of selective estrogen receptor modulators on colorectal cancer: a prognostic role for insulin-like growth factor binding protein-5

doi: 10.1042/CS20258451

Figure Lengend Snippet: Panels ( A , B ) illustrate the effect of varying concentrations of 27-OHC and E2 on the abundance of ERβ and IGFBP-5 in HCT-116 and HT-29 cell lines. The experiment was conducted in three biological repeats ( n = 3), with four different concentrations of 27-OHC (0.001, 0.1, 1, and 10 μM) or E2 (0.625, 1.25, 2.5, and 5 nM) used for treatment. The raw data from each Western blot experimental group were analysed through Image J and normalised against the phenol red-free SFM control group and were presented as fold changes relative to the control. Statistical analysis of differences was conducted using a one-way ANOVA with Dunnett's post hoc test showing with mean ± SD.

Article Snippet: This localization is consistent with data from the Human Protein Atlas, where IGFBP-5 is mainly detected in vesicles and is a secreted protein.

Techniques: Western Blot, Control

Journal: Clinical Science (London, England : 1979)

Article Title: Unravelling the effects of selective estrogen receptor modulators on colorectal cancer: a prognostic role for insulin-like growth factor binding protein-5

doi: 10.1042/CS20258451

Figure Lengend Snippet: ( A ) Proliferative responses of ERβ-silenced HCT-116 and HT-29 cells following 48-h treatment with 27-OHC (0.1and 1 μM) or E2 (5 nM). The experiment was conducted in three independent biological replicates, each performed in triplicate ( n = 3). Data are presented as mean ± SD and statistical analysis was performed using one-way ANOVA with Tukey multiple comparisons test. ( B , C ) Quantification of IGFBP-5, cPARP, and pH2AX protein levels in HCT-116 cells following 48-h treatment with 1 μM 27-OHC or 5 nM E2 in the presence or absence of ERβ knockdown. Protein expression was assessed by Western blotting. Densitometric analysis was performed using ImageJ software, and results are expressed as mean ± SD from three independent biological replicates ( n = 3), each carried out in triplicate. Statistical significance was evaluated using Student's t -test.

Article Snippet: This localization is consistent with data from the Human Protein Atlas, where IGFBP-5 is mainly detected in vesicles and is a secreted protein.

Techniques: Knockdown, Expressing, Western Blot, Software

Journal: Clinical Science (London, England : 1979)

Article Title: Unravelling the effects of selective estrogen receptor modulators on colorectal cancer: a prognostic role for insulin-like growth factor binding protein-5

doi: 10.1042/CS20258451

Figure Lengend Snippet: ( A ) Pearson correlation analysis between ERβ and IGFBP-5 mRNA expression levels in colon cancer and normal tissues using the GEPIA2 database. The x -axis represents IGFBP-5 mRNA expression, and the y -axis represents ERβ mRNA expression. A positive correlation suggests simultaneous increases or decreases in expression of both genes, whereas a negative correlation indicates inverse expression trends. The correlation coefficient ( R ) ranges from −1 to 1, with values closer to ±1 indicating a stronger correlation. A significant positive correlation was observed in colon cancer tissues, whereas a significant negative correlation was found in normal tissues. No statistically significant correlation was noted in rectal cancer tissues. ( B ) Relative mRNA expression of IGFBP-5 following ERβ silencing in HCT-116 and HT-29 colon cancer cell lines. The experiment was conducted in three independent biological replicates ( n = 3), and results are presented as mean ± SD. Statistical analysis was performed using Student's t -test. ( C ) Confocal immunofluorescence images showing the subcellular localization of ERβ (red) and IGFBP-5 (green) in HCT-116 and HT-29 cells, with nuclei counterstained using DAPI (blue). ERβ silencing visibly reduced IGFBP-5 signal intensity, supporting their regulatory association at the protein level. ( D ) Western blot analysis of IGFBP-5 protein expression in HCT-116 cells following ERβ knockdown. Densitometric quantification confirmed the down-regulation of IGFBP-5 protein in response to ERβ silencing. ( E ) Co-immunoprecipitation assay demonstrating the association between ERβ and IGFBP-5 in HCT-116 cells. ERβ was immunoprecipitated, and IGFBP-5 was detected in the precipitated complex by Western blotting, suggesting a potential physical interaction. ( F ) Protein–protein docking model of ERβ (pink) and IGFBP-5 (green) using ZDOCK 3.0.2, illustrating ten key interface residues at the predicted binding site. The top-ranked model based on ZDOCK scoring was selected for visualization. Structural analysis identified 24 residues within 3 Å, suggesting potential direct contact, as detailed in Supplementary Table S3.

Article Snippet: This localization is consistent with data from the Human Protein Atlas, where IGFBP-5 is mainly detected in vesicles and is a secreted protein.

Techniques: Expressing, Immunofluorescence, Western Blot, Knockdown, Co-Immunoprecipitation Assay, Immunoprecipitation, Binding Assay

Journal: Clinical Science (London, England : 1979)

Article Title: Unravelling the effects of selective estrogen receptor modulators on colorectal cancer: a prognostic role for insulin-like growth factor binding protein-5

doi: 10.1042/CS20258451

Figure Lengend Snippet: (A ) The effect of ERβ silencing on wound healing in HCT-116 colon cancer cells, as assessed by the HoloMonitor live-cell imaging system. Cells were transfected with ERβ siRNA and monitored for 24 h. Red-shaded areas represent regions of cell proliferation, whereas green-shaded areas indicate regions without cellular coverage. White regions reflect areas of high cell density that exceed the detection threshold of the laser-based imaging system. Compared with the control group, ERβ-silenced cells exhibited delayed migration and incomplete closure of the wound area, suggesting impaired migratory capacity following ERβ knockdown. ( B ) Differential expression of IGFBP-5 in CRC and matched normal tissues, analysed using the GEPIA2 platform. The box plots display gene expression in log2(TPM + 1) units, comparing tumour samples with normal samples from the TCGA and GTEx databases. For COAD, the analysis included 275 tumour and 349 normal samples; for rectum adenocarcinoma (READ), 92 tumour and 318 normal samples were analysed. Purple boxes represent normal tissues, and pink boxes indicate tumour tissues. Statistical comparisons were performed using Student's t -test. ( C ) Prognostic landscape of IGFBP-5 and ERs (including ERβ) in colon and rectal cancer, based on TCGA data. GEPIA2-generated risk maps show the prognostic value of gene expression, with red blocks denoting poor prognosis and blue blocks indicating favourable outcomes. Statistically significant results ( P <0.05) are outlined with a black frame. ( D ) Kaplan–Meier survival analysis based on TCGA colon adenocarcinoma database. The upper left panel shows overall survival (OS) stratified by IGFBP-5 expression. The upper right panel shows OS based on ERβ expression. The lower panel shows a two-gene survival analysis based on the combined expression of ERβ and IGFBP-5. The orange line represents high gene expression; the blue line indicates low expression. Censored data points are marked by plus signs (+). A steeper curve indicates a lower survival probability over time. ( E ) Pearson correlation analysis between IGFBP-5 and epithelial–mesenchymal transition (EMT)-related genes (CDH2/N-cadherin, VIM/vimentin, ZEB1, SOX2, Oct4, and Bmi1) in TCGA colon cancer tissues using GEPIA2. The x -axis represents IGFBP-5 mRNA expression levels, while the y -axis shows expression of individual EMT markers. Strong positive correlations were observed with N-cadherin, vimentin, and ZEB1.

Article Snippet: This localization is consistent with data from the Human Protein Atlas, where IGFBP-5 is mainly detected in vesicles and is a secreted protein.

Techniques: Live Cell Imaging, Transfection, Imaging, Control, Migration, Knockdown, Quantitative Proteomics, Gene Expression, Generated, Expressing

Journal: Clinical Science (London, England : 1979)

Article Title: Unravelling the effects of selective estrogen receptor modulators on colorectal cancer: a prognostic role for insulin-like growth factor binding protein-5

doi: 10.1042/CS20258451

Figure Lengend Snippet: A schematic to summarise the role of ERβ and GPER1 following exposure to 27-OHC, E2, or G1. The changing of ERβ and GPER1 leads to distinct downstream effects, including the down-regulation of IGFBP-5 and rapid non-genomic signalling, respectively. The dotted arrows indicate interactions or regulatory effects for which direct experimental evidence is currently lacking in the present study. Specifically, although silencing ERβ and treatment with E2 or 27-OHC results in down-regulation of IGFBP-5 and phenotypic alterations in CRC cells, a c ausal mechanistic link between ERβ activation and SERMs treatment has not been conclusively demonstrated. Moreover, the downstream effect of IGFBP-5 reduction on cell phenotypes is inferred based on correlation analysis and previous literature, but requires further validation. This schematic integrates current findings with working hypotheses to provide a conceptual framework for ERs signalling in CRC.

Article Snippet: This localization is consistent with data from the Human Protein Atlas, where IGFBP-5 is mainly detected in vesicles and is a secreted protein.

Techniques: Activation Assay, Biomarker Discovery

Journal: iScience

Article Title: Neuronal Igfbp2 deficiency in the prefrontal cortex impairs cognition through synaptic dysfunction in male mice

doi: 10.1016/j.isci.2026.115629

Figure Lengend Snippet: Neuron-specific Igfbp2 deficiency in the PFC induces cognitive dysfunction (A) Schematic of the experimental design. (B) Diagram of viral injection into the PFC (top) and representative image of viral expression (bottom); scale bars, 500 μm. (C) Representative western blot bands (top) and quantification of Igfbp2 protein levels in the PFC (bottom); n = 4 mice per group. (D) Schematic of the novel object recognition test, depicting the training phase (top) and the test phase (bottom). (E) Representative trajectory plots from the novel object recognition test. (F) Quantification of time spent investigating the novel object during the test phase; n = 8 mice per group. (G) Quantification of the number of investigations of the novel object during the test phase; n = 8 mice per group. (H) Schematic of the Y-maze test, depicting the training phase (top) and the test phase (bottom). (I) Representative trajectory plots from the Y-maze test. (J) Quantification of time spent in the novel arm during the test phase; n = 8 mice per group. (K) Quantification of the number of entries into the novel arm during the test phase; n = 8 mice per group. Analyzed by unpaired t test; ∗∗ p < 0.01. Data are presented as means ± SEM.

Article Snippet: Recombinant Igfbp2 protein (MCE; Cat# HY- P74846 ) was dissolved in sterile saline at 10 μg/μL.

Techniques: Injection, Expressing, Western Blot

Journal: iScience

Article Title: Neuronal Igfbp2 deficiency in the prefrontal cortex impairs cognition through synaptic dysfunction in male mice

doi: 10.1016/j.isci.2026.115629

Figure Lengend Snippet: Neuron-specific Igfbp2 deficiency in the PFC reduces excitatory synaptic transmission and neuronal excitability (A) Representative traces of sEPSC recorded from PFC neurons in AAV-scramble (top) and AAV-shRNA (bottom) groups. (B) Cumulative probability plots (main panel) and quantification (insert panel) of sEPSC frequency in AAV-Scramble and AAV-shRNA groups. (C) Cumulative probability plots (main panel) and quantification (insert panel) of sEPSC amplitude in AAV-Scramble and AAV-shRNA groups. (D) Representative traces of sIPSC recorded from PFC neurons in AAV-Scramble (top) and AAV-shRNA (bottom) groups. (E) Cumulative probability plots (main panel) and quantification (insert panel) of sIPSC frequency in AAV-Scramble and AAV-shRNA groups. (F) Cumulative probability plots (main panel) and quantification (insert panel) of sIPSC amplitude in AAV-Scramble and AAV-shRNA groups. (G) Representative traces of AP elicited by 200 pA current injections in PFC neurons from AAV-Scramble (left) and AAV-shRNA (right) groups. (H) Quantification of AP firing frequency in response to a range of current injections in PFC neurons from AAV-Scramble and AAV-shRNA groups. Analyzed by unpaired t test in B, C, E, and F, and two-way repeated-measure ANOVA in H; n = 8 neurons from 4 mice per group; ns: not significant, ∗∗ p < 0.01. Data are presented as means ± SEM.

Article Snippet: Recombinant Igfbp2 protein (MCE; Cat# HY- P74846 ) was dissolved in sterile saline at 10 μg/μL.

Techniques: Transmission Assay, shRNA

Journal: iScience

Article Title: Neuronal Igfbp2 deficiency in the prefrontal cortex impairs cognition through synaptic dysfunction in male mice

doi: 10.1016/j.isci.2026.115629

Figure Lengend Snippet: Neuron-specific Igfbp2 deficiency in the PFC disrupts synaptic integrity (A) Representative images of dendritic spines in the PFC from AAV-Scramble (left) and AAV-shRNA (right) groups; scale bars, 10 μm. (B) Quantification of dendritic spine density (number per 10 μm) in the PFC from AAV-Scramble and AAV-shRNA groups; n = 9 brain sections from 3 mice per group. (C) Representative western blot bands (top) and quantification of synapsin-1 protein levels (bottom) in the PFC from AAV-scramble and AAV-shRNA groups; n = 4 mice per group. (D) Representative western blot bands (top) and quantification of PSD-95 protein levels (bottom) in the PFC from AAV-Scramble and AAV-shRNA groups; n = 4 mice per group. Analyzed by unpaired t test; ∗ p < 0.05 and ∗∗∗ p < 0.001. Data are presented as means ± SEM.

Article Snippet: Recombinant Igfbp2 protein (MCE; Cat# HY- P74846 ) was dissolved in sterile saline at 10 μg/μL.

Techniques: shRNA, Western Blot

Journal: iScience

Article Title: Neuronal Igfbp2 deficiency in the prefrontal cortex impairs cognition through synaptic dysfunction in male mice

doi: 10.1016/j.isci.2026.115629

Figure Lengend Snippet: Exogenous supplementation of Igfbp2 in the PFC rescues cognitive deficits induced by neuron-specific Igfbp2 deficiency (A) Schematic of the experimental design. (B) Diagram shows virus injection and cannula placement into the PFC (top), and a representative image of cannula position and viral expression in the PFC (bottom); scale bars, 500 μm. (C) Representative western blot bands (top) and quantification of Igfbp2 protein levels in the PFC (bottom); n = 4 mice per group. (D) Schematic of the novel object recognition test, depicting the training phase (top) and the test phase (bottom). (E) Representative trajectory plots from the novel object recognition test. (F) Quantification of time spent investigating the novel object during the test phase; n = 8 mice per group. (G) Quantification of the number of investigations of the novel object during the test phase; n = 8 mice per group. (H) Schematic of the Y-maze test, depicting the training phase (top) and the test phase (bottom). (I) Representative trajectory plots from the Y-maze test. (J) Quantification of time spent in the novel arm during the test phase; n = 8 mice per group. (K) Quantification of the number of entries into the novel arm during the test phase; n = 8 mice per group. Analyzed by unpaired t test; ∗∗ p < 0.01. Data are presented as means ± SEM.

Article Snippet: Recombinant Igfbp2 protein (MCE; Cat# HY- P74846 ) was dissolved in sterile saline at 10 μg/μL.

Techniques: Virus, Injection, Expressing, Western Blot

Journal: iScience

Article Title: Neuronal Igfbp2 deficiency in the prefrontal cortex impairs cognition through synaptic dysfunction in male mice

doi: 10.1016/j.isci.2026.115629

Figure Lengend Snippet: Exogenous supplementation of Igfbp2 in the PFC restores excitatory synaptic transmission and neuronal excitability impaired by neuron-specific Igfbp2 deficiency (A) Representative traces of sEPSC recorded from PFC neurons in saline (top) and Igfbp2 (bottom) groups. (B) Cumulative probability plots (main panel) and quantification (insert panel) of sEPSC frequency in saline (top) and Igfbp2 (bottom) groups. (C) Cumulative probability plots (main panel) and quantification (insert panel) of sEPSC amplitude in saline and Igfbp2 groups. (D) Representative traces of sIPSC recorded from PFC neurons in saline (top) and Igfbp2 (bottom) groups. (E) Cumulative probability plots (main panel) and quantification (insert panel) of sIPSC frequency in saline and Igfbp2 groups. (F) Cumulative probability plots (main panel) and quantification (insert panel) of sIPSC amplitude in saline and Igfbp2 groups. (G) Representative traces of AP elicited by 200 pA current injections in PFC neurons from saline (left) and Igfbp2 (right) groups. (H) Quantification of AP firing frequency in response to a range of current injections in PFC neurons from Saline (left) and Igfbp2 (right) groups. Analyzed by unpaired t test in B, C, E, and F, and two-way repeated-measure ANOVA in H; n = 8 neurons from 4 mice per group; ns: not significant, ∗∗ p < 0.01. Data are presented as means ± SEM.

Article Snippet: Recombinant Igfbp2 protein (MCE; Cat# HY- P74846 ) was dissolved in sterile saline at 10 μg/μL.

Techniques: Transmission Assay, Saline

Journal: iScience

Article Title: Neuronal Igfbp2 deficiency in the prefrontal cortex impairs cognition through synaptic dysfunction in male mice

doi: 10.1016/j.isci.2026.115629

Figure Lengend Snippet: Exogenous supplementation of Igfbp2 in the PFC restores synaptic integrity disrupted by neuron-specific Igfbp2 deficiency (A) Representative images of dendritic spines in the PFC from saline (left) and Igfbp2 (right) groups; scale bars, 10 μm. (B) Quantification of dendritic spine density (number per 10 μm) in the PFC from saline and Igfbp2 groups; n = 9 brain sections from 3 mice per group. (C) Representative western blot bands (top) and quantification of synapsin-1 protein levels (bottom) in the PFC from AAV-Scramble and AAV-shRNA groups; n = 4 mice per group. (D) Representative western blot bands (top) and quantification of PSD-95 protein levels (bottom) in the PFC from AAV-Scramble and AAV-shRNA groups; n = 4 mice per group. Analyzed by unpaired t test; ∗ p < 0.05 and ∗∗∗ p < 0.001. Data are presented as means ± SEM.

Article Snippet: Recombinant Igfbp2 protein (MCE; Cat# HY- P74846 ) was dissolved in sterile saline at 10 μg/μL.

Techniques: Saline, Western Blot, shRNA

Journal: iScience

Article Title: Neuronal Igfbp2 deficiency in the prefrontal cortex impairs cognition through synaptic dysfunction in male mice

doi: 10.1016/j.isci.2026.115629

Figure Lengend Snippet: Schematic summary of this study Neuron-specific Igfbp2 deficiency in the PFC impairs cognitive function by disrupting synaptic integrity, excitatory transmission, and neuronal activity, whereas exogenous Igfbp2 supplementation mitigates cognitive deficits by restoring synaptic integrity, excitatory transmission, and neuronal activity. PFC prefrontal cortex, sEPSC spontaneous excitatory postsynaptic currents, Igfbp2 insulin-like growth factor-binding protein 2.

Article Snippet: Recombinant Igfbp2 protein (MCE; Cat# HY- P74846 ) was dissolved in sterile saline at 10 μg/μL.

Techniques: Transmission Assay, Activity Assay, Binding Assay