mab6434  (R&D Systems)

Journal: bioRxiv

Article Title: A systematic cross-modal approach identifies astrocytic VCAM1 as a regulator of hippocampal synapse development

doi: 10.64898/2026.03.17.712320

Figure Lengend Snippet: (A) Schematic of the workflow used to assess the effect of constitutive CRISPR/Cas9-mediated KO of Vcam1 or Gpr37l1 on hippocampal synapse development. SO: stratum oriens, SR: stratum radiatum, SLM: stratum lacunosum-moleculare, SL: stratum lucidum, DG: dentate gyrus, ML: molecular layer. Created with BioRender.com. (B) Quality control of Vcam1 KO by immunohistochemistry for VCAM1 and DAPI. Insets show CA1 region of both hemispheres. (C) Quality control of Gpr37l1 KO by immunohistochemistry for GPR37L1 and DAPI. Insets show CA1 region of both hemispheres. (D) Heatmap showing the log2 fold change in synaptic metrics comparing LacZ-gRNA and VCAM1-gRNA hemispheres across hippocampal laminae. For each synaptic metric and lamina, 3 FOVs (sections) per brain from 4 brains were analyzed. Log2-transformed metrics were analyzed using a linear mixed-effects model with gRNA, hippocampal lamina and their interaction as fixed effects to test gene- and lamina-specific effects, and brain as a random intercept to account for inter-brain variability. Asterisks denote significant differences after false discovery rate (FDR) correction for each metric across hippocampal laminae (FDR < 0.05). (E) Heatmap showing the log2 fold change in synaptic metrics comparing LacZ-gRNA and GPR37L1-gRNA hemispheres. Same sampling, plotting and analyses as in (G). (F) Representative processed Airyscan images of VGLUT1 and PSD95 in CA1 SR from LacZ-gRNA- and VCAM1-gRNA-injected hemispheres, with quantification of local peak max and presynapse MFI normalized to the LacZ-gRNA hemisphere. Each small data point represents a single FOV; larger linked data points represent hemisphere means. Statistical significance was assessed using a linear mixed effects model as in (G), followed by Tukey’s post hoc tests and FDR corrected significance. (G) Representative processed Airyscan images of VGLUT1 and PSD95 in CA3 SR from LacZ-gRNA- and VCAM1-gRNA-injected hemispheres, with quantification of local peak max and presynapse MFI normalized to the LacZ-gRNA hemisphere. Same sampling, plotting and analyses as in (F). (H) Schematic of the workflow used to assess the effect of VCAM1-Fc protein treatment on hippocampal synapses in vitro . Created with BioRender.com. (I) Representative images of hippocampal neurons treated with recombinant protein from DIV8 to DIV14. Fc or VCAM1-Fc (2 µg/mL) was added at DIV8 and replenished at DIV11. Neurons were fixed at DIV14 and stained for VGLUT1 (cyan), PSD95 (magenta), and MAP2 (yellow). Insets show the merge followed by the isolated VGLUT1-PSD95 signal. (J) Quantification of excitatory synaptic puncta density along MAP2+ dendrites. VGLUT1+PSD95+ colocalized puncta, as well as VGLUT1+ and PSD95+ puncta separately, were measured per 100 μm dendrite and normalized to the Fc condition. Each FOV contained two neuronal somas for standardized sampling; 9-11 FOVs per condition were analyzed in N = 4 independent experiments. Each data point represents one experiment is color-shaded by experiment. A linear mixed-effects model on log2 transformed data was used to account for inter-experimental variability. Post-hoc pairwise comparisons were conducted using Tukey’s HSD test. Individual FOV data points are shown in .

Article Snippet: Upon arrival, lyophilized recombinant mouse VCAM1-Fc (R&D Systems, 643-VM) was briefly centrifuged and reconstituted in sterile Dulbecco’s phosphate-buffered saline (DPBS).

Techniques: CRISPR, Control, Immunohistochemistry, Transformation Assay, Sampling, Injection, In Vitro, Recombinant, Staining, Isolation

Journal: bioRxiv

Article Title: A systematic cross-modal approach identifies astrocytic VCAM1 as a regulator of hippocampal synapse development

doi: 10.64898/2026.03.17.712320

Figure Lengend Snippet: (A) Western blot quality control of the recombinant proteins Fc and VCAM1-Fc used to treat hippocampal neurons, visualized using a total protein stain. (B) Representative images of hippocampal neurons treated with recombinant protein from DIV8 to DIV14. Fc or VCAM1–Fc (2 µg/mL) was added at DIV8 and replenished at DIV11. Neurons were fixed at DIV14 and stained for VGAT (cyan), GEPH (magenta), and MAP2 (yellow). Insets show the merge followed by the isolated VGAT-GEPH signal. (C) Quantification of inhibitory synaptic puncta density along MAP2+ dendrites. VGAT+GEPH+ colocalized puncta, as well as VGAT+ and GEPH+ puncta separately, were measured per 100 μm dendrite and normalized to Fc. Same sampling, plotting and analyses as in . (D) Plots from showing distribution of the analyzed FOVs. Each dot represents a single FOV and is color-shaded according to experiment. (E) Plots from (C) showing distribution of the analyzed FOVs. Each dot represents a single FOV and is color-shaded according to experiment. (F) Quantification of excitatory synaptic puncta size of VGLUT1+PSD95+ colocalized puncta, as well as VGLUT1+ and PSD95+ puncta separately. Each FOV contained two neuronal somas for standardized sampling; 9-11 FOVs per condition were analyzed in N = 4 independent experiments. Each data point represents one experiment (above panel), and one FOV (below panel), color-shaded by experiment. A linear mixed-effects model was used to account for multiple FOVs per experiment. (G) Quantification of inhibitory synaptic puncta size of VGAT+GEPH+ colocalized puncta, as well as VGAT+ and GEPH+ puncta separately. Same sampling, plotting and analyses as in (F).

Article Snippet: Upon arrival, lyophilized recombinant mouse VCAM1-Fc (R&D Systems, 643-VM) was briefly centrifuged and reconstituted in sterile Dulbecco’s phosphate-buffered saline (DPBS).

Techniques: Western Blot, Control, Recombinant, Staining, Isolation, Sampling

Journal: Cells

Article Title: Targeting Soluble VCAM1 and GSK3β Improves Cerebrovascular Function and Reduces Stroke Pathology in Diabetic Mice

doi: 10.3390/cells15050455

Figure Lengend Snippet: Glycemic control alone is insufficient to stabilize diabetic mast cells. Legends for the different treatment groups: Veh. Control-Vehicle control, LA-I-Long-acting Insulin, VmAb-VCAM1 monoclonal antibody, Tide.-Tideglusib. T2D refers to the H + S mice. ( A ) HOMA2-IR values calculated from glucose and insulin values of the respective treatment groups. n = 7 each, * p < 0.05 vs. Veh. control and # p < 0.05 vs. T2D (untreated). ( B ) Plasma histamine and ( C ) Plasma sVCAM1 levels in the different treatment groups. n = 7 each, * p < 0.05 vs. Veh. Control, # p < 0.05 vs. T2D (untreated), and δ p < 0.05 vs. T2D + LA-I alone.

Article Snippet: VmAb was an in vivo-grade, low-endotoxin, carrier-free anti-mouse VCAM1 antibody (#BE0027, BioXCell, Lebanon, NH, USA) administered intraperitoneally as a 15 mg/kg loading dose on day 1, followed by 10 mg/kg on days 4, 8, and 12.

Techniques: Control, Clinical Proteomics