rabbit anti lyve1  (Novus Biologicals)

Journal: Translational Stroke Research

Article Title: CircRNA circ_0004058 Modulates Early Brain Injury in Subarachnoid Hemorrhage Through miR-221-3p and VE1 Activation Pathway

doi: 10.1007/s12975-025-01383-9

Figure Lengend Snippet: Identification of key genes involved in SAH onset and validation in vivo. A : Volcano plot of differentially expressed genes between samples of brain tissue from SAH (n = 10) and sham (n = 10) rats according to RNA sequencing; red represents upregulated genes and green represents downregulated genes. B : Venn diagram of differentially expressed genes according to RNA sequencing and SAH-related genes analyzed by GeneCards. C–D : Expression levels of LYVE1 (C) and TEK (D) extracted from RNA sequencing data. E–F : mRNA and protein levels of LYVE1 in the hippocampal tissue of sham-operated and SAH rats, as detected by RT-qPCR (E) and Western blotting (F). G : Flow cytometry detection of LYVE1 + CD68 + macrophages in the hippocampal tissue of sham-operated and SAH rats. H–I : mRNA and protein levels of VEGF-A in the hippocampal tissue of sham-operated and SAH rats, as detected by RT-qPCR (H) and Western blotting (I). (J) Immunohistochemistry detection of CD31 and VEGFA protein levels in the hippocampal tissue of each group of rats. n = 8 rats per treatment. *** p < 0.001 vs. sham-operated rats

Article Snippet: After blocking with 5% bovine serum albumin, membranes were incubated overnight at 4 °C with rabbit monoclonal antibodies against LYVE1 (1:1000, #67,538, Cell Signaling Technology), vascular endothelial growth factor (VEGF)-A (1:1000, ab214424, Abcam), and GAPDH (ab181602, 1:10,000, Abcam).

Techniques: Biomarker Discovery, In Vivo, RNA Sequencing, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, Immunohistochemistry

Journal: Translational Stroke Research

Article Title: CircRNA circ_0004058 Modulates Early Brain Injury in Subarachnoid Hemorrhage Through miR-221-3p and VE1 Activation Pathway

doi: 10.1007/s12975-025-01383-9

Figure Lengend Snippet: Effects of LYVE1 overexpression on brain injury in SAH rats. ote: SAH rats were treated with oe-NC or oe-LYVE1. A : RT-qPCR detection of LYVE1 mRNA expression in the hippocampal tissue of SAH rats. B : Western blot of LYVE1 protein expression in the hippocampal tissue of SAH rats. C–D : Neurological function scores (C) and extent of cerebral edema (D) in SAH rats. E : The pathological scores of SAH rats. F : Representative images of brain specimens collected after SAH induction and quantitative assessment of SAH severity; G : Evans blue staining was used to evaluate BBB permeability and quantify Evans blue dye extravasation in each group; scale bar = 0.5 cm; H : Nissl staining of neuronal injury in the hippocampal tissue of SAH rats. I : TUNEL staining of neuronal apoptosis in the hippocampal tissue of SAH rats. J : Flow cytometry analysis of LYVE1 + CD68 + macrophages in the hippocampal tissue of SAH rats. K–L : mRNA and protein levels of VEGF-A in the hippocampal tissue of SAH rats, as detected by RT-qPCR (K) and Western blotting (L); M: Immunohistochemistry was used to detect CD31 and VEGFA protein levels in the hippocampal tissue of each group. n = 8 rats per treatment. ** p < 0.01, *** p < 0.001vs. treatment with oe-N

Article Snippet: After blocking with 5% bovine serum albumin, membranes were incubated overnight at 4 °C with rabbit monoclonal antibodies against LYVE1 (1:1000, #67,538, Cell Signaling Technology), vascular endothelial growth factor (VEGF)-A (1:1000, ab214424, Abcam), and GAPDH (ab181602, 1:10,000, Abcam).

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Western Blot, Staining, Permeability, TUNEL Assay, Flow Cytometry, Immunohistochemistry

Journal: Translational Stroke Research

Article Title: CircRNA circ_0004058 Modulates Early Brain Injury in Subarachnoid Hemorrhage Through miR-221-3p and VE1 Activation Pathway

doi: 10.1007/s12975-025-01383-9

Figure Lengend Snippet: Identification of miRNAs upstream of LYVE1. A : Volcano plot of differentially expressed miRNAs between control (n = 2) and SAH (n = 2) samples in the GSE161870 dataset. B: Venn diagram of regulatory miRNAs upstream of LYVE1, as predicted by miRWalk database, and differentially expressed miRNAs in the GSE161870 dataset. C : Heat map of the expression of candidate miRNAs in the GSE161870 dataset. D : miR-221-3p expression in the hippocampal tissue of sham-operated and SAH rats, as detected by RT-qPCR. E : miR-221-3p binding sites on LYVE1 mRNA, as predicted by miRWalk database. F : miR-221-3p binding to LYVE1, as confirmed by dual-luciferase reporter assay. G : Expression levels of miR-221-3p and LYVE1 in RMA-BMs co-cultured with primary rat hippocampal neurons that had been transfected with miR-221-3p mimic or miR-221-3p inhibitor and then treated with GW4869, as detected by RT-qPCR. H : Western blot of LYVE1 protein expression in RMA-BMs co-cultured with primary rat hippocampal neurons that had been transfected with miR-221-3p mimic or miR-221-3p inhibitor and then treated with GW4869. n = 8 rats per treatment. *** p < 0.001 vs. treatment with NC mimic or NC inhibitor. Cell experiments were repeated three times

Article Snippet: After blocking with 5% bovine serum albumin, membranes were incubated overnight at 4 °C with rabbit monoclonal antibodies against LYVE1 (1:1000, #67,538, Cell Signaling Technology), vascular endothelial growth factor (VEGF)-A (1:1000, ab214424, Abcam), and GAPDH (ab181602, 1:10,000, Abcam).

Techniques: Control, Expressing, Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Assay, Cell Culture, Transfection, Western Blot

Journal: Translational Stroke Research

Article Title: CircRNA circ_0004058 Modulates Early Brain Injury in Subarachnoid Hemorrhage Through miR-221-3p and VE1 Activation Pathway

doi: 10.1007/s12975-025-01383-9

Figure Lengend Snippet: Effects of miR-221-3p-mediated regulation of LYVE1 on EBI in SAH rats. Note: SAH rats were treated with miR-221-3p inhibitor alone or in combination with sh-LYVE1. A : RT-qPCR detection of miR-221-3p expression and LYVE1 mRNA expression in the hippocampal tissue of SAH rats. B : Western blot of LYVE1 protein expression in the hippocampal tissue of SAH rats. C – D : Neurological function scores (C) and extent of cerebral edema (D) in SAH rats. E : The pathological scores of SAH rats; F : Representative images of brain specimens collected after SAH induction and quantitative assessment of SAH severity. G : Evans blue staining to assess BBB permeability and quantify Evans blue dye extravasation in each group; scale bar = 0.5 cm; H : Nissl staining of neuronal injury in the hippocampal tissue of SAH rats. I : TUNEL staining of neuronal apoptosis in the hippocampal tissue of SAH rats. J : Flow cytometry analysis of LYVE1 + CD68. + macrophages in the hippocampal tissue of SAH rats. K–L : mRNA and protein levels of VEGF-A in the hippocampal tissue of SAH rats, as detected by RT-qPCR (K) and Western blotting (L). M : Immunohistochemistry to detect CD31 and VEGFA protein levels in the hippocampal tissue of each group. n = 8 rats per treatment. *** p < 0.001

Article Snippet: After blocking with 5% bovine serum albumin, membranes were incubated overnight at 4 °C with rabbit monoclonal antibodies against LYVE1 (1:1000, #67,538, Cell Signaling Technology), vascular endothelial growth factor (VEGF)-A (1:1000, ab214424, Abcam), and GAPDH (ab181602, 1:10,000, Abcam).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Staining, Permeability, TUNEL Assay, Flow Cytometry, Immunohistochemistry

Journal: Translational Stroke Research

Article Title: CircRNA circ_0004058 Modulates Early Brain Injury in Subarachnoid Hemorrhage Through miR-221-3p and VE1 Activation Pathway

doi: 10.1007/s12975-025-01383-9

Figure Lengend Snippet: Relationship among circ_0004058, miR-221-3p, and LYVE1. A : Volcano plot of differentially expressed circRNAs between control (n = 4) and SAH (n = 5) samples in the GSE161913 dataset. B : Venn diagram of regulatory circRNAs upstream of miR-221-3p, as predicted by circBank database, and differentially expressed circRNAs in the GSE161913 dataset. C : Differential expression analysis of circ_0000826 and circ_0004058 in the GSE161913 dataset (Control, n = 4; SAH, n = 5). D : Stability of circ_0004058, as determined by RNase R digestion. E : circ_0004058 expression in the hippocampal tissue of sham-operated and SAH rats, as detected by RT-qPCR. F : circ_0004058 and miR-221-3p binding sites, as predicted by starBase database. G : Binding of circ_0004058 to miR-221-3p, as confirmed by dual-luciferase reporter assay. H : circ_0004058 binding to miR-221-3p, as detected by RNA pull-down assays. I : circ_0004058 binding to miR-221-3p, assessed by RIP assays. J : Localization of circ_0004058 and miR-221-3p in primary rat hippocampal neurons, as detected by FISH assays. K : Expression levels of miR-221-3p and LYVE1 in RMA-BMs co-cultured with primary rat hippocampal neurons that had been transfected with oe-circ_0004058 or sh-circ_0004058 and then treated with GW4869, as detected by RT-qPCR. L : Western blot of LYVE1 protein expression in RMA-BMs co-cultured with primary rat hippocampal neurons that had been transfected with oe-circ_0004058 or sh-circ_0004058 and then treated with GW4869. n = 8 rats per treatment. ** p < 0.01, *** p < 0.001 vs. treatment with oe-NC or sh-NC. Cell experiments were repeated three times

Article Snippet: After blocking with 5% bovine serum albumin, membranes were incubated overnight at 4 °C with rabbit monoclonal antibodies against LYVE1 (1:1000, #67,538, Cell Signaling Technology), vascular endothelial growth factor (VEGF)-A (1:1000, ab214424, Abcam), and GAPDH (ab181602, 1:10,000, Abcam).

Techniques: Control, Quantitative Proteomics, Expressing, Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Assay, Cell Culture, Transfection, Western Blot

Journal: Translational Stroke Research

Article Title: CircRNA circ_0004058 Modulates Early Brain Injury in Subarachnoid Hemorrhage Through miR-221-3p and VE1 Activation Pathway

doi: 10.1007/s12975-025-01383-9

Figure Lengend Snippet: Effects of circ_0004058-mediated regulation of miR-221-3p and reduced EBI in SAH rats. Note: SAH rats were treated with oe-circ_0004058 alone or in combination with miR-221-3p mimic. A: RT-qPCR detection of circ_0004058, miR-221-3p, and LYVE1 expression in the hippocampal tissue of SAH rats. B: Western blot of LYVE1 protein expression in the hippocampal tissue of SAH rats. C–D: Neurological function scores (C) and extent of cerebral edema (D) in SAH rats. E: The pathological scores of SAH rats. F: Representative images of brain specimens collected after SAH induction and quantitative assessment of SAH severity. G: Evans blue staining to evaluate BBB permeability and quantitative analysis of Evans blue dye extravasation in each group; scale bar = 0.5 cm. H: Nissl staining of neuronal injury in the hippocampal tissue of SAH rats. I: TUNEL staining of neuronal apoptosis in the hippocampal tissue of SAH rats. J: Flow cytometry analysis of LYVE1 + CD68 + macrophages in the hippocampal tissue of SAH rats. K–L: mRNA and protein levels of VEGF-A in the hippocampal tissue of SAH rats, as detected by RT-qPCR (K) and Western blotting (L). M: Immunohistochemistry to detect CD31 and VEGFA protein levels in the hippocampal tissue of each group. n = 8 rats per treatment. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. treatment with oe-NC + NC mimic or oe-circ_0004058 + NC mimic

Article Snippet: After blocking with 5% bovine serum albumin, membranes were incubated overnight at 4 °C with rabbit monoclonal antibodies against LYVE1 (1:1000, #67,538, Cell Signaling Technology), vascular endothelial growth factor (VEGF)-A (1:1000, ab214424, Abcam), and GAPDH (ab181602, 1:10,000, Abcam).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Staining, Permeability, TUNEL Assay, Flow Cytometry, Immunohistochemistry

Journal: Translational Stroke Research

Article Title: CircRNA circ_0004058 Modulates Early Brain Injury in Subarachnoid Hemorrhage Through miR-221-3p and VE1 Activation Pathway

doi: 10.1007/s12975-025-01383-9

Figure Lengend Snippet: Effects of the circ_0004058/miR-221-3p/LYVE1 axis on EBI in SAH rats. Note: SAH rats were treated with oe-circ_0004058 alone or in combination with sh-LYVE1. A: RT-qPCR detection of circ_0004058, miR-221-3p, and LYVE1 expression in the hippocampal tissue of SAH rats. B: Western blot of LYVE1 protein expression in the hippocampal tissue of SAH rats. C–D: Neurological function scores (C) and extent of cerebral edema (D) in SAH rats. E: The pathological scores of SAH rats. F: Representative images of brain specimens collected after SAH induction and quantitative assessment of SAH severity. G: Evans blue staining to evaluate BBB permeability and quantitative analysis of Evans blue dye extravasation in each group; scale bar = 0.5 cm. H: Nissl staining of neuronal injury in the hippocampal tissue of SAH rats. I: TUNEL staining of neuronal apoptosis in the hippocampal tissue of SAH rats. J: Flow cytometry analysis of LYVE1 + CD68 + macrophages in the hippocampal tissue of SAH rats. K–L: mRNA and protein levels of VEGF-A in the hippocampal tissue of SAH rats, as detected by RT-qPCR (K) and Western blotting (L). M: Immunohistochemistry to detect CD31 and VEGFA protein levels in the hippocampal tissue of each group. n = 8 rats per treatment. *** p < 0.001 vs. treatment with oe-NC + sh-NC; # p < 0.05 vs. treatment with oe-circ_0004058 + sh-NC

Article Snippet: After blocking with 5% bovine serum albumin, membranes were incubated overnight at 4 °C with rabbit monoclonal antibodies against LYVE1 (1:1000, #67,538, Cell Signaling Technology), vascular endothelial growth factor (VEGF)-A (1:1000, ab214424, Abcam), and GAPDH (ab181602, 1:10,000, Abcam).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Staining, Permeability, TUNEL Assay, Flow Cytometry, Immunohistochemistry

Journal: Translational Stroke Research

Article Title: CircRNA circ_0004058 Modulates Early Brain Injury in Subarachnoid Hemorrhage Through miR-221-3p and VE1 Activation Pathway

doi: 10.1007/s12975-025-01383-9

Figure Lengend Snippet: Effects of LYVE1 + CD68 + macrophages on EBI in SAH rats. Note: SAH rats were treated with oe-LYVE1 alone or in combination with anti-CSF1R antibody. A: RT-qPCR detection of circ_0004058, miR-221-3p, and LYVE1 expression in the hippocampal tissue of SAH rats. B: Western blot of LYVE1 protein expression in the hippocampal tissue of SAH rats. C–D: Neurological function scores (C) and extent of cerebral edema (D) in SAH rats. E: The pathological scores of SAH rats. F: Representative images of brain specimens collected after SAH induction and quantitative assessment of SAH severity. G: Evans blue staining to assess BBB permeability and quantitative analysis of Evans blue dye extravasation in each group; scale bar = 0.5 cm. H: Nissl staining of neuronal injury in the hippocampal tissue of SAH rats. I: TUNEL staining of neuronal apoptosis in the hippocampal tissue of SAH rats. J: Flow cytometry analysis of LYVE1 + CD68 + macrophages in the hippocampal tissue of SAH rats. K–L: mRNA and protein levels of VEGF-A in the hippocampal tissue of SAH rats, as detected by RT-qPCR (K) and Western blotting (L). n = 8 rats per treatment. M: Immunohistochemistry to detect CD31 and VEGFA protein levels in the hippocampal tissue of each group. ** p < 0.01, *** p < 0.001 vs. treatment with oe-NC + anti-IgG; # p < 0.05 vs. treatment with oe-LYVE1 + anti-IgG

Article Snippet: After blocking with 5% bovine serum albumin, membranes were incubated overnight at 4 °C with rabbit monoclonal antibodies against LYVE1 (1:1000, #67,538, Cell Signaling Technology), vascular endothelial growth factor (VEGF)-A (1:1000, ab214424, Abcam), and GAPDH (ab181602, 1:10,000, Abcam).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Staining, Permeability, TUNEL Assay, Flow Cytometry, Immunohistochemistry

Journal: Translational Stroke Research

Article Title: CircRNA circ_0004058 Modulates Early Brain Injury in Subarachnoid Hemorrhage Through miR-221-3p and VE1 Activation Pathway

doi: 10.1007/s12975-025-01383-9

Figure Lengend Snippet: Schematic diagram of the mechanism by which the circ_0004058/miR-221-3p/LYVE1 axis affects SAH-induced EBI

Article Snippet: After blocking with 5% bovine serum albumin, membranes were incubated overnight at 4 °C with rabbit monoclonal antibodies against LYVE1 (1:1000, #67,538, Cell Signaling Technology), vascular endothelial growth factor (VEGF)-A (1:1000, ab214424, Abcam), and GAPDH (ab181602, 1:10,000, Abcam).

Techniques: