goat anti mouse cd31  (R&D Systems)

Journal: Communications Biology

Article Title: A perfusion-independent high-throughput method to isolate liver sinusoidal endothelial cells

doi: 10.1038/s42003-025-07458-5

Figure Lengend Snippet: a – c Mice were fed with a standard or CDAA diet for 10 weeks. a Images show Picrosirius Red staining on the liver sections. LSECs were isolated from healthy and fibrotic liver tissues. Scale bar = 100 µm. b The dot plot shows the number of LSECs isolated from fibrotic mouse liver. [mean ± SD, n = 4 mice]. c Quantitative PCR analyses to compare the expression level of various sinusoidal genes in LSECs isolated from either healthy or fibrotic mouse liver tissues. [mean ± SD, n = 4 mice]. * P < 0.05 (Mann–Whitney test). d – f Liver tissues collected from healthy pigs were processed to isolate LSECs. d FACS plot showing gating strategy to isolate CD31+ LSECs and CD31− cells. e The dot plot shows the number of LSECs isolated from pig liver tissues. [mean ± SD, n = 4 pigs]. f Quantitative PCR analyses to compare expression of various vascular genes between CD31+ LSECs and CD31− cells. [mean ± SD, n = 4 pigs]. * P < 0.05 (Mann–Whitney test).

Article Snippet: Cells were subsequently washed with PBS, fixed in 4% PFA [Carl Roth, Cat #0335.1], blocked with 10% normal donkey serum [Biozol, Cat #LIN-END9010-10] and incubated in a primary antibody mix consisting of different combinations of goat anti-mouse CD31 [R&D Systems, Cat #AF3628], goat anti-mouse CD32b [R&D Systems, Cat #AF1460], rabbit anti-mouse Desmin [Abcam, Cat # ab15200], rabbit anti-mouse ERG [Abcam, Cat #ab196149], rat anti-mouse F4/80 [BioLeegend, # 123102], and rabbit anti-mouse Stabilin-2 antibodies overnight.

Techniques: Staining, Isolation, Real-time Polymerase Chain Reaction, Expressing, MANN-WHITNEY

Journal: Communications Biology

Article Title: A perfusion-independent high-throughput method to isolate liver sinusoidal endothelial cells

doi: 10.1038/s42003-025-07458-5

Figure Lengend Snippet: a – d LSECs were isolated using perfusion independent digestion method, followed by positive selection using CD146-magnetic beads. After that, enriched LSECs were placed in culture. a Primary LSEC cultures were stained for an endothelial cell marker (ERG, in green) and nuclear marker (DAPI, in gray). On the left, images show IF-stained LSEC cultures. Scale bar = 25 µm. The yellow arrow highlights an ERG-negative cell. On the right, the dot plot shows the percentage of ERG-positive cells of DAPI-positive cells. [mean ± SD, n = 3 wells]. b Images show IF-stained LSECs with CD31 (in gray), Stabilin-2 (in green), and DAPI (in blue). Scale bar = 25 µm. c Images show IF-stained LSECs with CD32b (in gray) and ERG (in green). Scale bar = 25 µm. d Representative scanning electron microscopy images of cultured LSECs. Scale bar = 2 µm.

Article Snippet: Cells were subsequently washed with PBS, fixed in 4% PFA [Carl Roth, Cat #0335.1], blocked with 10% normal donkey serum [Biozol, Cat #LIN-END9010-10] and incubated in a primary antibody mix consisting of different combinations of goat anti-mouse CD31 [R&D Systems, Cat #AF3628], goat anti-mouse CD32b [R&D Systems, Cat #AF1460], rabbit anti-mouse Desmin [Abcam, Cat # ab15200], rabbit anti-mouse ERG [Abcam, Cat #ab196149], rat anti-mouse F4/80 [BioLeegend, # 123102], and rabbit anti-mouse Stabilin-2 antibodies overnight.

Techniques: Isolation, Selection, Magnetic Beads, Staining, Marker, Electron Microscopy, Cell Culture

Journal: Communications Biology

Article Title: A perfusion-independent high-throughput method to isolate liver sinusoidal endothelial cells

doi: 10.1038/s42003-025-07458-5

Figure Lengend Snippet:

Article Snippet: Cells were subsequently washed with PBS, fixed in 4% PFA [Carl Roth, Cat #0335.1], blocked with 10% normal donkey serum [Biozol, Cat #LIN-END9010-10] and incubated in a primary antibody mix consisting of different combinations of goat anti-mouse CD31 [R&D Systems, Cat #AF3628], goat anti-mouse CD32b [R&D Systems, Cat #AF1460], rabbit anti-mouse Desmin [Abcam, Cat # ab15200], rabbit anti-mouse ERG [Abcam, Cat #ab196149], rat anti-mouse F4/80 [BioLeegend, # 123102], and rabbit anti-mouse Stabilin-2 antibodies overnight.

Techniques:

Journal: Scientific Reports

Article Title: Peripheral nerve regeneration using a bioresorbable silk fibroin-based artificial nerve conduit fabricated via a novel freeze–thaw process

doi: 10.1038/s41598-025-88221-y

Figure Lengend Snippet: Promotion of vascularization and macrophage-related immune response after transplantation. The SFC group showed a higher degree of vascularization and M2 macrophage recruitment than the other groups. ( A ) Representative images of the NC, SFC, Auto, and Sham groups observed using light microscopy, showing the area around the regenerated nerves, immunostained for CD31. ( B ) The total CD31-positive area of the SFC group was greater than that of the other groups, indicating that the SFC promoted vascularization inside the conduit. ( C ) Representative images of the NC, SFC, Auto, and Sham groups observed using light microscopy, showing the area around and inside the regenerated nerves, immunostained for Iba1 and arginase-1. (D) The total Iba-1 and arginase-1 double-positive area of the SFC group was greater than that of the other groups, indicating that the SFC promoted M2 macrophage recruitment.

Article Snippet: After blocking with the blocking solution; Blocking One (Nacalai Tesque, Kyoto, Japan), 20 times diluted with 0.1 M PBS containing 0.2% TritonX-100, the following primary antibodies were applied: rabbit polyclonal anti-neurofilament heavy (NFH) polypeptide antibody (ab8135; 1:500 dilution; Abcam), chicken myelin basic protein polyclonal anti-peptide antibody (MBP88837983; 1:500 dilution; Aves), goat polyclonal anti-mouse and anti-rat CD31 (AF3628; 1:100 dilution; R&D), rabbit polyclonal anti-Iba1 antibody (GtX100042; 1:500 dilution; GeneTex), and goat polyclonal anti-arginase-1 antibody (ab60176; 1:100 dilution; Abcam).

Techniques: Transplantation Assay, Light Microscopy

Journal: Angiogenesis

Article Title: Neuropilin-1 controls vascular permeability through juxtacrine regulation of endothelial adherens junctions

doi: 10.1007/s10456-024-09963-3

Figure Lengend Snippet: NRP1 regulates VEGFA-mediated permeability in an organotypic manner. A Schematic illustration of experimental design to induce recombination in Nrp1 fl/fl ; Cdh5 CreERT2 ( Nrp1 iECKO) mice. Nrp1 fl/fl were used as control. B and C RNA in-situ hybridisation (ISH) analysis of Nrp1 expression in the ear dermis ( B ) and back skin ( C ) of Nrp1 iECKO mice. Images highlight Nrp1 mRNA particles (arrows) specific to CD31-positive vessel area and graphs show quantification of Nrp1 mRNA expression ( Nrp1 mRNA particle area/vessel area). Scale bar: 50 μm. D Representative images showing leakage of 2000 kDa FITC-dextran (Pseudo-colour) in response to intradermal VEGFA injection in ear dermis of control and Nrp1 iECKO mice. E Leakage sites per vessel length in response to intradermal VEGFA stimulation in the ear skin of control and Nrp1 iECKO mice. n = 6 mice, two or more acquisitions/mouse. F Quantification of 2000 kDa dextran extravasation over time in the ear skin of control and Nrp1 iECKO mice following intradermal VEGFA stimulation. Black lines represent lines of best fit for the slope between leakage initiation and leakage termination. N ≥ 3 mice, two or more acquisitions/mouse, three or more sites/acquisition. G Evans blue leakage following intradermal administration of VEGFA in the ear skin of control and Nrp1 iECKO mice. Top, representative images. Bottom, quantification of Evans blue extravasation shown as VEGFA-induced leakage fold over PBS control (n ≥ 4 mice). H and I Leakage of fixable 2000 kDa FITC dextran in back skin ( H ) and trachea ( I ) after systemic administration of VEGFA in control and Nrp1 iECKO mice Left, representative images. Right, quantification of tracer leakage area/vessel area (n ≥ 8 mice, 2 or more fields of view/mouse). J Evans blue leakage with intradermal administration of VEGFA in the back skin of control and Nrp1 iECKO mice. Top, representative images. Bottom, quantification of Evans blue extravasation shown as VEGFA-induced leakage fold over PBS control (n ≥ 4 mice). Error bars; mean ± SEM. Statistical significance: Two-tailed unpaired Student’s t-test and linear regression with ANCOVA. Scale bar: 100 μm unless stated

Article Snippet: Commercial antibodies used were: rat anti-CD31 (BD Biosciences, 553370), goat anti-CD31 (R&D Systems, AF3628), goat anti-NRP1 (R&D Systems, AF566), chicken anti-GFP (Abcam, Ab13970), rabbit anti-phospho-PLCγ1 (Tyr783) (Cell signaling technology, 2821s), rabbit anti-PLCγ1 (Cell signaling technology, 2822s), and goat anti–VE-cadherin (R&D systems, AF1002).

Techniques: Permeability, Control, In Situ, Hybridization, Expressing, Injection, Two Tailed Test

Journal: Acta Histochemica et Cytochemica

Article Title: Localization of Both CD31- and Endomucin-Expressing Vessels in Mouse Dental Pulp

doi: 10.1267/ahc.24-00009

Figure Lengend Snippet: Localization of CD31 (+) and endomucin (+) vessels. ( A ) Stage of tooth crown formation at 1 week. Endomucin (+) vessels (red) mainly exist in the dental pulp. ( B ) Higher magnifications of the boxed regions in A . Both CD31(+) and endomucin (+) vessels (yellow) are narrow and located beneath the coronal dentin. ( C ) Higher magnifications of the boxed regions in B . Merged immunoreaction (yellow) is localized at thin vessels in the odontoblast layer. ( D ) Dental pulp at 4 weeks. Both CD31 (+) and endomucin (+) vessels decreased in the crown and increased in the root of dental pulp at 4 weeks. ( E ) Higher magnifications of the boxed regions in D . Thin vessels are colored in yellow, and express both CD31 (+) and endomucin (+) and located near the radicular dentin (arrowheads). ( F ) Higher magnifications of the boxed regions in E . Note the merged immunoreaction (yellow) shows mesh-like pattern. ( G ) Dental pulp at 12 weeks. Root canals at 12 week are narrower than those at 4 week. ( H ) Higher magnifications of the boxed regions in G . CD31 (+)-immunoreaction is mainly seen, and Both CD31 (+) and endomucin (+) vessels are faintly seen in dental pulp (arrowhead). ( I ) Higher magnifications of the boxed regions in H . CD31 (+) vessels are seen beneath the odontoblast layer. ( J ) Dental pulp at 56 weeks. Few CD31 (+) and endomucin (+) vessels were detected in dental pulp. ( K ) Higher magnifications of the boxed regions in J . Main vessels are positive for CD31 in the coronal pulp. ( L ) Higher magnifications of the boxed regions in K . Immuno-reaction for endomucin was separately seen at the CD31(+) vessels. Green: CD31 (+), Red: Endomucin (+), Yellow: CD31 (+) and endomucin (+). Arrowheads: CD31 (+) and endomucin (+) vessels. Den: dentin; Ob: odontoblast layer. Bars = 200 μm ( A , D , G , J ), 50 μm ( B , E , H , K ) and 15 μm ( C , F , I , L ).

Article Snippet: Sections of decalcified samples were incubated with an Alexa Fluor 488-conjugated anti-CD31/PECAM-1 goat polyclonal antibody (dilution 1:100, FAB3628G-10, R&D Systems, Minneapolis, MN, USA) and a rat monoclonal antibody against mouse endomucin (dilution 1:200, sc-65495, Santa Cruz Biotechnology Inc., Dallas, TX, USA) at 20 ± 5°C for 8 hr or 4°C overnight in the dark.

Techniques:

Journal: Acta Histochemica et Cytochemica

Article Title: Localization of Both CD31- and Endomucin-Expressing Vessels in Mouse Dental Pulp

doi: 10.1267/ahc.24-00009

Figure Lengend Snippet: Ratio of CD31 (+) and endomucin (+) vessels. Data represent means ± standard deviations (SD). Four mice were used for statistical analysis in each group. These ratios 12 and 56 weeks were significantly lower than those 1 and 4 weeks. * Statistical significance ( P < 0.05)

Article Snippet: Sections of decalcified samples were incubated with an Alexa Fluor 488-conjugated anti-CD31/PECAM-1 goat polyclonal antibody (dilution 1:100, FAB3628G-10, R&D Systems, Minneapolis, MN, USA) and a rat monoclonal antibody against mouse endomucin (dilution 1:200, sc-65495, Santa Cruz Biotechnology Inc., Dallas, TX, USA) at 20 ± 5°C for 8 hr or 4°C overnight in the dark.

Techniques:

Journal: Acta Histochemica et Cytochemica

Article Title: Localization of Both CD31- and Endomucin-Expressing Vessels in Mouse Dental Pulp

doi: 10.1267/ahc.24-00009

Figure Lengend Snippet: Calcein labeling and endomucin (+) vessels. ( A ) At 4 weeks after birth. Dentin with a large distance between the two calcein-lines was observed in the crown and root dentin. ( B ) Higher magnification of the boxed regions in A . Both CD31(+) and endomucin (+) vessels colored in white were thin and present directly beneath calcein-labeled dentin at all sites. ( C ) At 8 weeks after birth. Although the lines were present in root dentin, they were negligible in coronal dentin. The distance between the two lines at 8 weeks were narrower than that at 4 weeks. Both CD31 and endomucin (+) vessels were scattered in coronal dental pulp. ( D ) Higher magnification of the boxed regions in C . Both CD31 and endomucin (+) vessels were located at the odontoblast layer in root pulp. ( E ) At 12 weeks after birth. Calcein-labeled areas were only present in newly formed apical cementum, and were faint or not seen at coronal and root dentin. ( F ) Higher magnification of the boxed regions in E . CD31(+) and ndomucin (+) vessels were seen neat the newly formed cementum and faintly detected in dental pulp. Den: dentin, Cem: cementum, Ob: odontoblast layer, Pu: dental pulp. Bars = 200 μm ( A , C , E ), 30 μm ( B , D , F ). Green: calcein labeling, Cyan: CD31(+) vessel, Red: endomucin (+) vessel, White: both CD31(+) and endomucin(+) vessel, Blue: nuclear.

Article Snippet: Sections of decalcified samples were incubated with an Alexa Fluor 488-conjugated anti-CD31/PECAM-1 goat polyclonal antibody (dilution 1:100, FAB3628G-10, R&D Systems, Minneapolis, MN, USA) and a rat monoclonal antibody against mouse endomucin (dilution 1:200, sc-65495, Santa Cruz Biotechnology Inc., Dallas, TX, USA) at 20 ± 5°C for 8 hr or 4°C overnight in the dark.

Techniques: Labeling