mouse anti aqp4  (R&D Systems)

Journal: The Journal of Clinical Investigation

Article Title: Neutrophil-microglia interaction drives motor dysfunction in a neuromyelitis optica model induced by subarachnoid AQP4-IgG

doi: 10.1172/JCI199706

Figure Lengend Snippet: ( A ) Experimental design of the IgG binding to AQP4 on astrocytes in primary glial cultures established from wild-type and Aqp4 –/– pup brains. Microglia were depleted by treating Clodrosome (100 μg/mL) before subculture. Two days after adding IgG or cytokines, CXCL1 production was assessed by immunostaining or ELISA. ( B ) Immunoblot analysis of WT, Aqp4 –/– primary glial cells using IgGs specific for AQP4 and actin (loading control). ( C ) CXCL1 immunoreactivity was assessed in wild-type and AQP4 ± cells exposed to a control nonpathogenic monoclonal mouse IgG specific for the AQP4 cytoplasmic C-terminal domain (CCD-AQP4-IgG, m5) or a pathogenic extracellular domain–reactive IgG (ECD-AQP4-IgG, m21) or to IFN-γ plus TNF-α cytokines. Astrocytes are identified by AQP4 and GFAP immunoreactivities; microglia by IBA1; DNA is blue (DAPI). ( D ) Cellular CXCL1 (upper) was quantified from fluorescence intensity in C images; secreted CXCL1 protein levels (lower) were quantified in the glial culture media by ELISA. ( E ) Experimental design: 2 groups of WT mice were infused continuously via spinal subarachnoid catheter (day 0 to day 7) with pathogenic ECD-AQP4-IgG or ECD-AQP4-IgG mixed with CXCL1-IgG. ( F ) Motor function of mice in E was assessed by rotarod test; time: F (8, 40) = 1.319, P = 0.2632; treatment: F (1, 5) = 16.38, P = 0.0098; interaction: F (8, 22) = 7.766, P < 0.0001; n = 4–5 mice per group. ( G and H ) Representative images at day 7 of IgG infusion and quantification of neutrophils (Ly6G + ) in lumbar cord parenchyma ( n = 4 mice per group). Higher magnifications (left corners) of boxed areas in G show neutrophils (magenta). One-way ANOVA with Tukey’s post hoc in D ; n = 4 wells in D ; 2-way repeated measures ANOVA with Holm-Šídák post hoc test in F ; t test in H .

Article Snippet: After 3 days’ training on the rotarod beam (days –3 to –1, 4 rpm/min, 5 min) with daily slow manual catheter flushing with artificial CSF, an osmotic minipump delivery system (ALZET, 1007D), containing a pathogenic mouse monoclonal anti-AQP4-IgG1 (clone mECD, isotype, IgG1κ, AQP4-m21, created in-house; licensed to Sigma-Aldrich as MABN2471) or mouse polyclonal serum IgG (Sigma, I5381), or anti-AQP4-IgG plus rat monoclonal anti-CXCL1-IgG (R&D Systems, MAB453, clone 48415, 0.5 μg/12 μL/day for 8 days), was placed subcutaneously between the shoulders.

Techniques: Binding Assay, Immunostaining, Enzyme-linked Immunosorbent Assay, Western Blot, Control, Fluorescence

Journal: bioRxiv

Article Title: Conserved phenotype and function of human brain border-associated macrophages in iPSC-derived models

doi: 10.64898/2025.12.11.693582

Figure Lengend Snippet: ( A ) Schematic diagram of experimental design for generating iPS-derived HPCs for transplantation (see Methods ). ( B ) Example image of human xBAM (labeled with anti-human CD163) located along a pial vessel (labeled with anti-mouse CD31) in the leptomeninges (delineated with anti-mouse AQP4).

Article Snippet: The following antibodies were used in this study: anti-Iba1 (rabbit, Wako; 1:500); anti-human Nuclei (mouse, 3E1.3 Milipor MAB4383; 1:100); anti-mouse CD31 (goat, R&D AF3628; 1:100), anti-human CD163 (mouse, Invitrogen EDHu-1, MA1-82342; 1:500), anti-human Tmem119 (rabbit, Abcam AB185333; 1:100); anti-mouse AQP4 (rabbit, Novus Bio, NBP1-87679; 1:100).

Techniques: Derivative Assay, Transplantation Assay, Labeling

Journal: bioRxiv

Article Title: Conserved phenotype and function of human brain border-associated macrophages in iPSC-derived models

doi: 10.64898/2025.12.11.693582

Figure Lengend Snippet: ( A ) Schematic diagram of experimental approach for generating human-murine chimeric brain immune compartment. ( B and C ) Whole-section tile scan demonstrating distribution of human-derived cells in chimeric brain and zoomed image demonstrating co-habitation of both human (magenta nuclei) and murine macrophages (yellow arrows). ( D ) Whole section tile scan of distribution of human CD163 + cells in chimeric brain. Yellow arrows point to example cells localized in leptomeninges. ( E ) Zoomed image demonstrating co-habitation of both human microglia-like cells (xMGs; TMEM119 + CD163 - ) and human BAM-like cells (xBAMs; TMEM119 - CD163 + ; yellow arrows). ( F and G ) Confocal image and 3-dimensional rendering of xBAM along brain vasculature. Human CD163 + xBAM is located beyond glia limitans as evidenced by AQP4 stain. See also .

Article Snippet: The following antibodies were used in this study: anti-Iba1 (rabbit, Wako; 1:500); anti-human Nuclei (mouse, 3E1.3 Milipor MAB4383; 1:100); anti-mouse CD31 (goat, R&D AF3628; 1:100), anti-human CD163 (mouse, Invitrogen EDHu-1, MA1-82342; 1:500), anti-human Tmem119 (rabbit, Abcam AB185333; 1:100); anti-mouse AQP4 (rabbit, Novus Bio, NBP1-87679; 1:100).

Techniques: Derivative Assay, Staining

Journal: The European Journal of Neuroscience

Article Title: VEGF‐E Attenuates Injury After Ischemic Stroke by Promoting Reparative Revascularization

doi: 10.1111/ejn.70114

Figure Lengend Snippet: VEGF‐E‐mediated revascularization promotes astrocyte endfeet association. (a) Representative fluorescence images of CD31and AQP4 immunolabeling in the ipsilateral brain in vehicle (VEH)‐ and VEGF‐E‐treated mice 4 days after stroke. (b) Analysis of the density of perivascular AQP4 + structures in the ipsilateral hemisphere. (c) Analysis of CD31 and AQP4 colocalization in the ipsilateral hemisphere. Data are boxplot with min/max or stacked histogram ( n = 6–7 animals/group). ** p < 0.01 compared to control (b, unpaired two‐tailed t ‐test). Abbreviations: IF, immunofluorescence; VEGF, vascular endothelial growth factor; VEH, vehicle.

Article Snippet: The following primary antibodies were used: rat anti‐mouse cluster of differentiation (CD31) (1/500; BD Biosciences, ON, Canada; 550274), goat anti‐mouse aminopeptidase N (CD13) (1/250; R&D systems), rat anti‐mouse glial fibrillary acidic protein (GFAP) (1/500; Invitrogen, MA, USA; 13‐0300), rabbit anti‐mouse ionized calcium binding adaptor molecule (IBA1) (1/500; WAKO, ON, Canada; 019‐19741), goat anti‐rat CD45 (1/500; BD Biosciences; 553076), rabbit anti‐mouse PDGFRβ (1/250; Abcam, ON, Canada; ab32570), rat anti‐mouse endoglin (CD105) (1/250; Novus Biologicals, ON, Canada; NB100‐77666), and rabbit anti‐mouse aquaporine‐4 (AQP4) (1/500; Cell Signaling Technology, MA, USA; 59678S).

Techniques: Fluorescence, Immunolabeling, Control, Two Tailed Test, Immunofluorescence

Journal: The European journal of neuroscience

Article Title: VEGF-E Attenuates Injury After Ischemic Stroke by Promoting Reparative Revascularization.

doi: 10.1111/ejn.70114

Figure Lengend Snippet: FIGURE 4 | VEGF-E-mediated revascularization promotes astrocyte endfeet association. (a) Representative fluorescence images of CD31and AQP4 immunolabeling in the ipsilateral brain in vehicle (VEH)- and VEGF-E-treated mice 4 days after stroke. (b) Analysis of the density of perivas- cular AQP4+ structures in the ipsilateral hemisphere. (c) Analysis of CD31 and AQP4 colocalization in the ipsilateral hemisphere. Data are boxplot with min/max or stacked histogram (n = 6–7 animals/group). **p < 0.01 compared to control (b, unpaired two-tailed t-test). Abbreviations: IF, immu- nofluorescence; VEGF, vascular endothelial growth factor; VEH, vehicle.

Article Snippet: The following primary antibodies were used: rat anti- mouse cluster of differentiation (CD31) (1/500; BD Biosciences, ON, Canada; 550274), goat anti- mouse aminopeptidase N (CD13) (1/250; R&D systems), rat anti- mouse glial fibrillary acidic protein (GFAP) (1/500; Invitrogen, MA, USA; 13- 0300), rabbit anti- mouse ionized calcium binding adaptor molecule (IBA1) (1/500; WAKO, ON, Canada; 019- 19741), goat anti- rat CD45 (1/500; BD Biosciences; 553076), rabbit anti- mouse PDGFRβ (1/250; Abcam, ON, Canada; ab32570), rat anti- mouse endoglin (CD105) (1/250; Novus Biologicals, ON, Canada; NB100- 77666), and rabbit anti- mouse aquaporine- 4 (AQP4) (1/500; Cell Signaling Technology, MA, USA; 59678S).

Techniques: Fluorescence, Immunolabeling, Control, Two Tailed Test