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

Journal: bioRxiv

Article Title: Circulating biomarkers in serum from aortic valve stenosis patients predict sex-specific drug responses in valve myofibroblasts

doi: 10.64898/2026.01.16.700010

Figure Lengend Snippet: Schematic overview of the approach to link candidate serum biomarkers with in vitro drug efficacy. B, αSMA gradient mean intensity for male and female VICs cultured on hydrogels with sex-matched AVS patient serum (N=16 male, N=15 female). Statistical significance is indicated as *** = P < 0.001 (unpaired two-tailed t-test with Welch’s correction). C, Percent αSMA reduction in male and female VICs cultured on hydrogels with optimized doses of Ataciguat, Colchicine, or Evogliptin. Statistical significance is indicated as ** = P < 0.01 (unpaired two-tailed t-test with Welch’s correction). D, Heatmaps showing the correlation between each candidate serum factor and antifibrotic drug efficacy for male or female VICs cultured with sex-matched AVS serum (N=16 male, N=15 female). Statistical significance is indicated as * = P < 0.05 (paired two-tailed t-test). E, Correlation between serum IGFBP2 abundance (relative fluorescence units (RFU)) and percent αSMA reduction in male and female VICs cultured on gels with sex-matched AVS serum and Evogliptin (N=16 male, N=15 female).

Article Snippet: Human IGFBP2 neutralizing antibody (R&D systems, Cat. No AF674) and normal IgG control antibody (R&D systems, Cat. No AB-108-C) were resuspended in sterile PBS at 200 μg/mL and 1,000 μg/mL respectively.

Techniques: In Vitro, Cell Culture, Two Tailed Test, Fluorescence

Journal: bioRxiv

Article Title: Circulating biomarkers in serum from aortic valve stenosis patients predict sex-specific drug responses in valve myofibroblasts

doi: 10.64898/2026.01.16.700010

Figure Lengend Snippet: IGFBP2 mediates Evogliptin resistance in female VICs cultured with female AVS serum . A, Serum IGFBP2 concentrations quantified using an ELISA kit (N=31 AVS, N=5 healthy). Statistical significance is indicated as **** = P < 0.0001 (unpaired two-tailed t-test with Welch’s correction). B through C, Percent αSMA reduction in (B) female and (C) male VICs cultured on hydrogels with FBS and treated with Evogliptin and/or recombinant human IGFBP2 protein (N=6 gels). Statistical significance is indicated as * = P < 0.05 (one sample t and Wilcoxon test against a theoretical mean of zero). D, Serum IGFBP2 levels quantified using an ELISA kit with and without IGFBP2 neutralizing antibody (IGFBP2 Ab), IgG control antibody, and recombinant human IGFBP2 protein (N=3 male, N=3 female). Statistical significance is indicated as *** = P < 0.001 (one-way ANOVA with Tukey posttests). E, Percent αSMA reduction in male and female VICs cultured on hydrogels with sex-matched AVS patient serum samples, Evogliptin, IgG, IGFBP2 Ab, or recombinant human IGFBP2 protein (N=3 male, N=3 female). Statistical significance is indicated as *** = P < 0.001 (one-way ANOVA with Tukey posttests). F, Representative immunofluorescent images of male and female VICs cultured on hydrogels (green: αSMA, blue: nuclei). Scale bar = 100 μm. G, Normalized IGFBP to IGF1 ratio for male and female AVS serum samples (N=23 male, N=16 female). Statistical significance is indicated as *** = P < 0.001 (unpaired two-tailed t-test with Welch’s correction). H, Correlation between normalized IGFBP to IGF1 ratios and percent αSMA reduction in male and female VICs cultured on hydrogels with sex-matched serum and Evogliptin.

Article Snippet: Human IGFBP2 neutralizing antibody (R&D systems, Cat. No AF674) and normal IgG control antibody (R&D systems, Cat. No AB-108-C) were resuspended in sterile PBS at 200 μg/mL and 1,000 μg/mL respectively.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Recombinant, Control

Journal: bioRxiv

Article Title: Circulating biomarkers in serum from aortic valve stenosis patients predict sex-specific drug responses in valve myofibroblasts

doi: 10.64898/2026.01.16.700010

Figure Lengend Snippet: Principal component analysis clustering samples based on RNA sequencing results (N=3). B through C, Volcano plot of differential gene expression from RNA sequencing between (B) Vehicle and Evogliptin groups or (C) Vehicle and Evogliptin plus IGFBP2 neutralizing antibody groups (N=3). Statistical significance was defined as FDR-adjusted P-value < 0.05 (unpaired two-tailed t-test with Welch’s correction) and fold change greater than 2. D, Venn diagram comparing differentially expressed genes in the Evogliptin and Evogliptin plus IGFBP2 neutralizing antibody groups relative to vehicle control (N=3). E, Volcano plot of differential gene expression from RNA sequencing between Evogliptin and Evogliptin plus IGFBP2 neutralizing antibody groups (N=3). Statistical significance was defined as FDR-adjusted P-value < 0.05 (unpaired t-test with Welch’s correction) and fold change greater than 2. F, Heatmap of the top 20 up and down regulated genes in the Evogliptin plus IGFBP2 neutralizing antibody group relative to vehicle control (N=3). G through H, Top 15 upregulated pathways in Evogliptin versus Evogliptin plus IGFBP2 neutralizing antibody group identified by (G) KEGG and (H) Ingenuity Pathway Analysis. Abbreviations: Evo: Evogliptin, Evo + Ab: Evogliptin with IGFBP2 neutralizing antibody, Veh: Vehicle control. Number indicates patient serum sample ID (26, 30, and 45).

Article Snippet: Human IGFBP2 neutralizing antibody (R&D systems, Cat. No AF674) and normal IgG control antibody (R&D systems, Cat. No AB-108-C) were resuspended in sterile PBS at 200 μg/mL and 1,000 μg/mL respectively.

Techniques: RNA Sequencing, Gene Expression, Two Tailed Test, Control

Journal: bioRxiv

Article Title: Circulating biomarkers in serum from aortic valve stenosis patients predict sex-specific drug responses in valve myofibroblasts

doi: 10.64898/2026.01.16.700010

Figure Lengend Snippet: IGFBP2 promotes Evogliptin resistance in female VICs cultured with female AVS serum through Rho/ROCK and focal adhesion kinase signaling. A, Schematic overview of the approach to validate the mechanism of IGFBP2-mediated Evogliptin resistance. B, Percent αSMA reduction in female VICs cultured on hydrogels with female AVS serum and treated with various doses of H-1152 with and without Evogliptin (N=3 gels). For H-1152: + = 0.1 μM; ++ = 0.5 μM; +++ = 5 μM. Statistical significance is indicated as * = P < 0.05 (one-way ANOVA with Tukey posttests). C, Percent αSMA reduction in female VICs cultured on hydrogels with AVS serum and treated with various doses of PF-573228 with and without Evogliptin (N=3 gels). For PF-573228: + = 0.1 μM; ++ = 1 μM; +++ = 10 μM. Statistical significance is indicated as * = P < 0.05 (one-way ANOVA with Tukey posttests). D through E, Percent αSMA reduction in (D) female or (E) male VICs cultured on hydrogels with sex-matched AVS serum and treated with various combinations of Evogliptin, IGFBP2 neutralizing antibody, H-1152, and PF-573228 (N=3 serum samples per sex). Statistical significance is indicated as * = P < 0.05, ** = P < 0.01 (one-way ANOVA with Tukey posttests). F, Representative immunofluorescent images of male and female VICs cultured on hydrogels with sex-matched serum and various drug combinations (green: αSMA, blue: nuclei). Scale bar = 100 μm. Abbreviations: Evo = Evogliptin, H11 = H-1152, PF5 = PF-573228, Ab = IGFBP2 neutralizing antibody.

Article Snippet: Human IGFBP2 neutralizing antibody (R&D systems, Cat. No AF674) and normal IgG control antibody (R&D systems, Cat. No AB-108-C) were resuspended in sterile PBS at 200 μg/mL and 1,000 μg/mL respectively.

Techniques: Cell Culture

Journal: bioRxiv

Article Title: Circulating biomarkers in serum from aortic valve stenosis patients predict sex-specific drug responses in valve myofibroblasts

doi: 10.64898/2026.01.16.700010

Figure Lengend Snippet: Summary mechanistic figure illustrating how elevated IGFBP2 in female serum modulates Evogliptin efficacy in female VICs.

Article Snippet: Human IGFBP2 neutralizing antibody (R&D systems, Cat. No AF674) and normal IgG control antibody (R&D systems, Cat. No AB-108-C) were resuspended in sterile PBS at 200 μg/mL and 1,000 μg/mL respectively.

Techniques: