rat anti cd13  (Bio-Rad)

Journal: bioRxiv

Article Title: Blockade of VCAM1 or VLA4 preserves cerebrovasculature and prevents cognitive decline late after stroke

doi: 10.1101/2025.06.25.661593

Figure Lengend Snippet: A) Total vessel length in the peri-infarct cortex after 3 weeks of chronic isotype control (Stroke-α-IC) or anti-VCAM1 (Stroke-α-VCAM1) antibody treatment, using endothelial staining for CD31 to define vasculature. B) Representative confocal images and (C) quantification of pericyte (CD13+) coverage of CD31+ vasculature in the peri-infarct cortex or contralateral cortical region of stroked mice treated with isotype control (Stroke-α-IC) or anti-VCAM1 (Stroke-α-VCAM1) antibody. D) Total vessel length in the peri-infarct cortex after 3 weeks of chronic isotype control (Stroke-α-IC) or anti-VLA4 (Stroke-α-VLA4) antibody treatment. E) Representative confocal images and (F) quantification of pericyte (CD13+) coverage of CD31+ vasculature in the peri-infarct cortex or contralateral cortical region of stroked mice treated with isotype control (Stroke-α-IC) or anti-VLA4 (Stroke-α-VLA4) antibody. Scale bar, 20 µM; Statistics, Student’s t test or 2-way ANOVA with Tukey’s post hoc test for group comparisons; Error bars, mean ± SEM; *p < 0.05; **p < 0.01.

Article Snippet: Primary antibodies were used against CD13 (1:500; Bio-Rad, MCA2183GA), CD31 (1:200; Novus Biologicals, AF3628 or 1:300, BD Biosciences, 550274), fibrinogen (1:1000; DAKO, A0080), and GFAP (1:1000; Abcam, Ab4674).

Techniques: Control, Staining

Journal: bioRxiv

Article Title: Reactive Pericytes Lead to Microvascular Dysfunction and Cortical Neurodegeneration During Experimental Autoimmune Encephalomyelitis

doi: 10.1101/2025.02.17.638582

Figure Lengend Snippet: A. Immunofluorescence staining reveals pericyte coverage (CD13, red) and endothelial proliferating cells marked by CD105 (vasculature, green) in the cortex of control and induced EAE mice. Representative confocal images were taken using 20X objectives and magnification was applied with 40X in both control and EAE mice. B. Quantification of vascular pericyte coverage reveals a %103 increase in the cortex of induced EAE mice compared to control mice (***p≤0.001). C. Quantification of the number of pericyte cell bodies on the microvasculature demonstrates no significant difference in the cortex of induced EAE mice compared to control mice ( ns p=0.36). Statistical analysis was performed using a two-tailed unpaired t-test, with n=3 in the control group and n=6 in the EAE group. Error bars represent the mean with +/− SEM. Scale bars in all images represent 50µm.

Article Snippet: The following primary antibodies were used: Rat anti-Mouse CD13 (1:100; Bio-Rad; MCA2183), Rabbit anti-Mouse CD105 (1:200; Abcam; ab221675), Rat anti-Mouse CD31 (1:200; BD Pharmingen; 550274), Dylight 488-conjugated tomato-lectin (1:300; ThermoFisher; DL-1174), Rabbit anti-NeuN (1:200; Abcam; ab177487), Rat anti-Mouse CD45 (1:200; BioLegend; 103102), and Rabbit anti-Neurofilament heavy polypeptide (1:200; Abcam; ab207176).

Techniques: Immunofluorescence, Staining, Control, Two Tailed Test

Journal: bioRxiv

Article Title: Reactive Pericytes Lead to Microvascular Dysfunction and Cortical Neurodegeneration During Experimental Autoimmune Encephalomyelitis

doi: 10.1101/2025.02.17.638582

Figure Lengend Snippet: A. Immunofluorescent staining reveals pericyte coverage (CD13, red) and endothelial proliferating cells marked by CD105 (vasculature, green) in the cortex of control and induced EAE mice. Representative confocal images were taken using 20X objectives in both control and EAE mice. B. Quantification of vascular pericyte coverage reveals a %118 increase in the hippocampus of induced EAE mice compared to control mice (**p=0.002). C. Quantification of the number of pericyte cell bodies on the microvasculature demonstrates no significant difference in the hippocampus of induced EAE mice compared to control mice (ns p=0.31). Statistical analysis was performed using a two-tailed unpaired t-test, with n=3 in the control group and n=4 in the EAE group. Error bars represent the median with interquartile range. Scale bars in all images represent 50µm.

Article Snippet: The following primary antibodies were used: Rat anti-Mouse CD13 (1:100; Bio-Rad; MCA2183), Rabbit anti-Mouse CD105 (1:200; Abcam; ab221675), Rat anti-Mouse CD31 (1:200; BD Pharmingen; 550274), Dylight 488-conjugated tomato-lectin (1:300; ThermoFisher; DL-1174), Rabbit anti-NeuN (1:200; Abcam; ab177487), Rat anti-Mouse CD45 (1:200; BioLegend; 103102), and Rabbit anti-Neurofilament heavy polypeptide (1:200; Abcam; ab207176).

Techniques: Staining, Control, Two Tailed Test

Journal: bioRxiv

Article Title: Reactive Pericytes Lead to Microvascular Dysfunction and Cortical Neurodegeneration During Experimental Autoimmune Encephalomyelitis

doi: 10.1101/2025.02.17.638582

Figure Lengend Snippet: A. Confocal microscopy images show plasma-derived IgG stained (cyan), along with CD105-stained vessels (green) and CD13-stained pericytes (red), in the cortex of control and induced EAE mice. Images were taken by 20X objectives. B. Quantification of plasma-derived IgG in the cortex reveals a substantial ∼ 161% (over 16-fold) increase in luminal IgG deposition in induced EAE mice compared to control mice (*p= 0.04). C. Plasma-derived IgG shows significant an increase in the luminal region of the microvasculature, accompanied by increased pericyte coverage in the same area. Statistical analysis was performed using a two-tailed unpaired t-test (n=3 in the control group and n=4 in the EAE group). Error bar represents mean with +/− SEM. Scale bars are 100 µm in A and 10µm in B, respectively.

Article Snippet: The following primary antibodies were used: Rat anti-Mouse CD13 (1:100; Bio-Rad; MCA2183), Rabbit anti-Mouse CD105 (1:200; Abcam; ab221675), Rat anti-Mouse CD31 (1:200; BD Pharmingen; 550274), Dylight 488-conjugated tomato-lectin (1:300; ThermoFisher; DL-1174), Rabbit anti-NeuN (1:200; Abcam; ab177487), Rat anti-Mouse CD45 (1:200; BioLegend; 103102), and Rabbit anti-Neurofilament heavy polypeptide (1:200; Abcam; ab207176).

Techniques: Confocal Microscopy, Derivative Assay, Staining, Control, Two Tailed Test

Journal: bioRxiv

Article Title: Reactive Pericytes Lead to Microvascular Dysfunction and Cortical Neurodegeneration During Experimental Autoimmune Encephalomyelitis

doi: 10.1101/2025.02.17.638582

Figure Lengend Snippet: A. Confocal microscopy images show plasma-derived IgG stained (cyan), along with CD105-stained vessels (green) and CD13-stained pericytes (red), in the hippocampus of control and induced EAE mice. Images were taken by 20X objectives. B. Quantification of plasma-derived IgG in the hippocampus reveals a substantial increase in luminal IgG deposition in induced EAE mice compared to control mice (*p= 0.04). C. with meticulous observation, plasma-derived IgG shows deposition in both focal and diffuse area within the cortex and hippocampus, specifically in the luminal region of the microvasculature. Statistical analysis was performed using a two-tailed unpaired t-test, n=3 in the control group and n=4 in the EAE group. Error bar represents median with interquartile range. Scale bars are 50µm.

Article Snippet: The following primary antibodies were used: Rat anti-Mouse CD13 (1:100; Bio-Rad; MCA2183), Rabbit anti-Mouse CD105 (1:200; Abcam; ab221675), Rat anti-Mouse CD31 (1:200; BD Pharmingen; 550274), Dylight 488-conjugated tomato-lectin (1:300; ThermoFisher; DL-1174), Rabbit anti-NeuN (1:200; Abcam; ab177487), Rat anti-Mouse CD45 (1:200; BioLegend; 103102), and Rabbit anti-Neurofilament heavy polypeptide (1:200; Abcam; ab207176).

Techniques: Confocal Microscopy, Derivative Assay, Staining, Control, Two Tailed Test

Journal: bioRxiv

Article Title: Reactive Pericytes Lead to Microvascular Dysfunction and Cortical Neurodegeneration During Experimental Autoimmune Encephalomyelitis

doi: 10.1101/2025.02.17.638582

Figure Lengend Snippet: Captured immunofluorescent images were indicated staining of CD105 (green), CD13 (red), CD45+ cells (cyan) and Hoechst nuclear staining (blue) in the cortex of induced EAE mice. 20X magnification for control and EAE groups and 40X for the higher magnification were used in EAE mice. Scale bars representing 20µm.

Article Snippet: The following primary antibodies were used: Rat anti-Mouse CD13 (1:100; Bio-Rad; MCA2183), Rabbit anti-Mouse CD105 (1:200; Abcam; ab221675), Rat anti-Mouse CD31 (1:200; BD Pharmingen; 550274), Dylight 488-conjugated tomato-lectin (1:300; ThermoFisher; DL-1174), Rabbit anti-NeuN (1:200; Abcam; ab177487), Rat anti-Mouse CD45 (1:200; BioLegend; 103102), and Rabbit anti-Neurofilament heavy polypeptide (1:200; Abcam; ab207176).

Techniques: Staining, Control

Journal: bioRxiv

Article Title: Reactive Pericytes Lead to Microvascular Dysfunction and Cortical Neurodegeneration During Experimental Autoimmune Encephalomyelitis

doi: 10.1101/2025.02.17.638582

Figure Lengend Snippet: A. Immunofluorescent staining of CD105 (green) and CD45+ cells (cyan) reveals increased intravascular stalling (yellow arrowheads) and rare trafficking of CD45+ leukocytes (yellow arrow) through the microvasculature in the EAE mice, accompanied by increased or stalling red blood cells within the intravascular compartment, as indicated by the red arrow. Images were taken with 20X objective and magnification was performed with 40X in both the control and EAE mice. B, C. Quantification results indicate a significant increase in intravascular and extravasated CD45+ cells in EAE-induced mice (*p=0.03, *p=0.03, respectively). The statistical analysis was performed using a two-tailed unpaired t-test (n=3 in the control and n=4 in the EAE mice). Error bar represented mean with +/− SEM and scale bars are 20µm. D. Immunofluorescent staining of the cortical vasculature highlights microglial activation, marked by IBA1 (green), and platelet endothelial adhesion molecules, marked by CD31 (red), in both control and EAE-induced mice. Images were taken using 20X objective. E. Quantification of IBA1 intensity in the cortex reveals a significant increase in microglial activation in induced EAE mice compared to control mice (*p=0.04). Statistical analysis was performed using two-tailed unpaired t-test with n=3 in the control and n=6 in the EAE mice. Error bar represented mean with +/− SEM and scale bars are 100µm. F. Immunofluorescent staining of CD45+ cells (cyan), CD105 (green), CD13 (red) and Hoechst (blue) reveals that CD45 was also expressed on activated microglia under inflammatory conditions in the EAE compared to the control mice. The scale bars are representing of 50µm.

Article Snippet: The following primary antibodies were used: Rat anti-Mouse CD13 (1:100; Bio-Rad; MCA2183), Rabbit anti-Mouse CD105 (1:200; Abcam; ab221675), Rat anti-Mouse CD31 (1:200; BD Pharmingen; 550274), Dylight 488-conjugated tomato-lectin (1:300; ThermoFisher; DL-1174), Rabbit anti-NeuN (1:200; Abcam; ab177487), Rat anti-Mouse CD45 (1:200; BioLegend; 103102), and Rabbit anti-Neurofilament heavy polypeptide (1:200; Abcam; ab207176).

Techniques: Staining, Control, Two Tailed Test, Activation Assay

Journal: bioRxiv

Article Title: Reactive Pericytes Lead to Microvascular Dysfunction and Cortical Neurodegeneration During Experimental Autoimmune Encephalomyelitis

doi: 10.1101/2025.02.17.638582

Figure Lengend Snippet: Representative confocal images show immunofluorescent staining of CD105 (green), CD13 (red), CD45+ cells (cyan) and Hoechst nuclear staining (blue) in the cortex of induced EAE mice. The images were captured using 20X and 40X objectives, with scale bars representing 50µm.

Article Snippet: The following primary antibodies were used: Rat anti-Mouse CD13 (1:100; Bio-Rad; MCA2183), Rabbit anti-Mouse CD105 (1:200; Abcam; ab221675), Rat anti-Mouse CD31 (1:200; BD Pharmingen; 550274), Dylight 488-conjugated tomato-lectin (1:300; ThermoFisher; DL-1174), Rabbit anti-NeuN (1:200; Abcam; ab177487), Rat anti-Mouse CD45 (1:200; BioLegend; 103102), and Rabbit anti-Neurofilament heavy polypeptide (1:200; Abcam; ab207176).

Techniques: Staining

Journal: bioRxiv

Article Title: Reactive Pericytes Lead to Microvascular Dysfunction and Cortical Neurodegeneration During Experimental Autoimmune Encephalomyelitis

doi: 10.1101/2025.02.17.638582

Figure Lengend Snippet: Representative confocal images of the spinal cord (top row) and cortex (bottom row) indicate immunoflourecent staining of activated microglial with tomato lectin (green), CD13 (red), and Hoechst nuclear staining (blue) in both spinal cord and cortex of the induced EAE mice. The images were captured using 40X objectives, with scale bars representing 50µm.

Article Snippet: The following primary antibodies were used: Rat anti-Mouse CD13 (1:100; Bio-Rad; MCA2183), Rabbit anti-Mouse CD105 (1:200; Abcam; ab221675), Rat anti-Mouse CD31 (1:200; BD Pharmingen; 550274), Dylight 488-conjugated tomato-lectin (1:300; ThermoFisher; DL-1174), Rabbit anti-NeuN (1:200; Abcam; ab177487), Rat anti-Mouse CD45 (1:200; BioLegend; 103102), and Rabbit anti-Neurofilament heavy polypeptide (1:200; Abcam; ab207176).

Techniques: Staining