dylighttm 594 vector laboratories dl  (Vector Laboratories)

Journal: bioRxiv

Article Title: Endothelial-pericyte interactions regulate angiogenesis via VEGFR2 signaling during retinal development and disease

doi: 10.1101/2025.03.08.642174

Figure Lengend Snippet: ( A ) Schematic illustration of experimental schedule in the developing mouse. ( B ) Left - Representative immunofluorescence images of the healthy mouse retinal tissue at P17 throughout different layers. Right - Quantification of the presence of pericytes (NG2, green) relative to mouse vasculature (Isolectin B4, red) (n=6). Scale bar: 50µm. ( C ) Conclusion schematic showing that downregulation of VEGFR2 through direct contact between ECs and pericytes enhances pericyte recruitment, thereby leading to the stabilization of the nascent vasculature. Illustrations created with BioRender.com.

Article Snippet: Immunofluorescence staining was conducted by permeabilizing and blocking the cryosections with 5% normal goat serum containing 0.05% TWEEN 20 in PBS follow by incubation of DyLight 594-conjugated Griffonia Simplicifolia Lectin I (GSL I) Isolectin B4 (Vector Laboratories) overnight at 4°C in the blocking buffer.

Techniques: Immunofluorescence

Journal: bioRxiv

Article Title: Endothelial-pericyte interactions regulate angiogenesis via VEGFR2 signaling during retinal development and disease

doi: 10.1101/2025.03.08.642174

Figure Lengend Snippet: ( A ) Schematic illustration of experimental schedule of OIR mouse model. ( B ) Representative western blots for VEGFR2 pY951 and beta-actin in mouse retina collated 24 hours after ZM323881 injection. ( C ) i. Representative immunofluorescence images of the OIR mouse retinal tissue at P17, five days after intravitreal injection of VEGFR2 inhibitor (Isolectin B4, red; VO area, blue; NV area, white). ii-iii. Quantifications of vaso-obliteration area (VO) and pathological neovascularization (NV) at P17 (n=15). Scale bar: 300 µm. ( D ) Left - Representative immunofluorescence images of the OIR mouse retinal tissue at P17 throughout different layers. Right - Quantification of the presence of pericytes (NG2, green) relative to mouse vasculature (Isolectin B4, red) (n=11). Scale bar: 50 µm. ( E ) i. Representative immunofluorescence images of the OIR mouse retina cross-section at P17. ( ii-iii ) Quantification of retina thickness and the percentage of vessel coverage (Isolectin B4, red) relative to retina area (DAPI, blue) (n=3 mice, 8 sections each). Scale bar: 100 µm. ( F ) i . Schematic illustration of experimental schedule of vascular permeability assay with OIR mouse model. ii. Representative immunofluorescence images of the OIR mouse retinal tissue (left). Scale bar: 50 µm. iii. Relative fluorescence intensity of FITC-dextran in OIR retinas treated with or without VEGFR2 inhibitor. Bars represent means±SD; n=6. Illustrations created with BioRender.com. Bars represent means±SD; Significance levels were set at not significant (ns) p > 0.05, *p ≤ 0.05, ***p ≤ 0.001 and ****p ≤ 0.0001.

Article Snippet: Immunofluorescence staining was conducted by permeabilizing and blocking the cryosections with 5% normal goat serum containing 0.05% TWEEN 20 in PBS follow by incubation of DyLight 594-conjugated Griffonia Simplicifolia Lectin I (GSL I) Isolectin B4 (Vector Laboratories) overnight at 4°C in the blocking buffer.

Techniques: Western Blot, Injection, Immunofluorescence, Permeability, Fluorescence