polyclonal rabbit anti mouse lyve 1  (AngioBio Inc)

Journal: medRxiv

Article Title: Intracalvariosseous bone marrow injection detours the blood-brain barrier as a novel drug delivery approach for pre-clinic and clinic trials in stroke

doi: 10.1101/2024.03.03.24303548

Figure Lengend Snippet: a. Process of injecting Cy3-NA1 by ICO, 647-CD31 by intravenous and 488-Lyve1 by intracisternal magna. b. Skull-dura-brain integrated tissue processing timeline. c. Overall scan results of lymphatic vessels, blood vessels and Cy3-NA1, scale=2 mm. d. Cy3-NA1 enters brain tissue through microchannels (white arrows) between the skull and brain parenchyma after ICO injection, scale=1 mm, inset=300 μm.

Article Snippet: After injecting 5 ul of Cy3-labeled NA1 (Cy3-NA1) into the mice’s skull bone marrow, 10 ul of 488-labeled Rabbit polyclonal Lyve1 antibody (R&D, FAB2125G) was immediately injected through the cisterna magna at a speed of 2 ul/min.

Techniques: Injection

Journal: PLoS ONE

Article Title: CXCR4 Is Dispensable for T Cell Egress from Chronically Inflamed Skin via the Afferent Lymph

doi: 10.1371/journal.pone.0095626

Figure Lengend Snippet: Immunofluorescence staining of frozen sections of chronically inflamed skin (21 days after inflammation elicited by subcutaneous injection of CFA) for CXC12 and CCL21 by LYVE-1 + lymph vessels using DAPI counterstaining. One representative image of lymph vessels analyzed in 5 mice is shown. Scale bars, 10 µm.

Article Snippet: The slides were then incubated with polyclonal rabbit anti-mouse LYVE-1 (AngioBio) and goat anti-mouse CCL21 (R&D Systems) or monoclonal mouse anti-mouse CXCL12 (clone 79018; R&D Systems).

Techniques: Immunofluorescence, Staining, Injection

Journal: Drug Design, Development and Therapy

Article Title: Inhibition of lymphangiogenesis in vitro and in vivo by the multikinase inhibitor nintedanib

doi: 10.2147/DDDT.S130297

Figure Lengend Snippet: Systemic application of BIBF 1120 (nintedanib) significantly decreased angiogenesis and hemangiogenesis in the cornea in comparison with the controls. Notes: ( A1 – A3 ) Non-sutured corneas were used as negative controls. Sutured corneas with nintedanib treatment ( C1 – C3 ) were compared to those without nintedanib treatment ( B1 – B3 ). Significant inhibition of ( D ) lymphangiogenesis ( P <0.01, n=8) and ( E ) hemangiogenesis ( P <0.01, n=8) after systemic treatment with nintedanib over 14 days was observed compared with the controls without nintedanib treatment. A–C at 100× magnification. Abbreviation: LYVE-1, lymphatic vessel endothelial hyaluronan receptor-1.

Article Snippet: Mice were sacrificed at planned times, and the eyes were removed and dissected; following this, whole-mounted corneas were fixed in 4% paraformaldehyde overnight at 4°C and blocked in 5% donkey serum albumin (Solarbio, Beijing, China) for 1 h. For double lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) and CD31 immunostainings, corneas were incubated overnight with polyclonal rabbit anti-mouse LYVE-1 (1/200; Abcam, Cambridge, UK) and rat anti-mouse CD31 (1/100; BD Biosciences Pharmingen, San Jose, CA, USA) antibodies, as described previously.

Techniques: Inhibition

Journal: bioRxiv

Article Title: Multi-omics Analysis Identifies IgG2b Class-Switching with ALCAM-CD6 Co-Stimulation in Lymph Nodes During Advanced Inflammatory-Erosive Arthritis

doi: 10.1101/2022.10.27.514103

Figure Lengend Snippet: PLNs from WT (5.5-month-old; n=10 PLNs) (A) and TNF-Tg mice with Early (5-6-month-old; n=6 PLNs) (B) and Advanced (>8-month-old; n=12 PLNs, 6/capture area) (C-D) arthritis were harvested and processed for spatial transcriptomics. Each H&E-stained image (left) corresponds to a transcriptional representation (right) of the PLN sinus (red spots overlaying white sinus spaces), paracortical (green spots), and follicular (blue spots) regions. Isolated spots with high ribosomal gene content were annotated as an undefined (purple) cell population (A-E) . Feature plots represent the identification of Lyve Hi /Marco Hi sinuses (F-G) , Ptprc Hi paracortices (H) , and Ptprc Hi /Ms4a1 Hi follicles (I) that were validated by histologic localization as in A-D . Unsupervised clustering (resolution=0.5) resolved 11 transcriptionally distinct PLN regions, where the sinus-associated spots localized to 3 distinct clusters (dashed lines) (J) . The Ighg1 Hi / Ighg2b Hi / Ighg2c Hi / Ighm Hi Sinus-Immuno (K-N) and Lyve1 Hi / Marco Hi Sinus-LEC (F-G) populations predominated, while Sinus-Undefined spots exhibited a high proportion of ribosomal genes. The predominant Sinus-Immuno (red) and Sinus-LEC (blue) populations were subset and re-clustered (O) , and individual PLNs were evaluated for total sinus-associated spots (Sinus-Immuno + Sinus-LEC) (P) and proportion of Sinus-Immuno spots (Q) . Statistics: One-way ANOVA, *p<0 . 05, **p<0 . 01 (P , Q) .

Article Snippet: Antibodies and dilutions for immunofluorescent staining: Rabbit anti-mouse CD6 (ThermoFisher, Cat# MA5-29680, RRID: AB_2785505, 1:50), rat anti-mouse F4/80 (BioRad, Cat# MCA497R, RRID:AB_323279, 1:50), rabbit anti-mouse Marco (Abcam, Cat# ab239369, 1:100), Alexa Fluor 555 goat anti-mouse IgG2b cross-adsorbed (ThermoFisher Scientific, Cat# A-21147, RRID:AB_2535783, 1:50), APC rat anti-mouse CD45R/B220 (BD Biosciences, Cat# 553092, RRID:AB_398531, 1:100), goat anti-mouse Alcam (CD166; ThermoFisher, Cat# PA5-47083, RRID:AB_2607383, 1:100), Cy3 goat anti-mouse IgM (Jackson ImmunoResearch Laboratories, Cat# 115-165-020, RRID:AB_2338683, 1:200), FITC donkey anti-mouse IgG (Jackson ImmunoResearch Laboratories, Cat# 715-095-150, RRID:AB_2340792, 1:200), goat anti-proliferating cell nuclear antigen (PCNA; Santa Cruz Biotechnology, Cat# sc-9857, RRID:AB_2160372, 1:50), FITC Peanut Agglutinin (PNA; Millipore Sigma, L7381-1MG, 1:50), goat anti-mouse peripheral node addressin (PNAd; BD Biosciences, Cat#553863, RRID:AB_395099, 1:50), rabbit anti-mouse Lyve1 (Acris Antibodies, Cat# DP3513, RRID:AB_1004776, 1:50), AlexaFluor 568 donkey anti-goat IgG (ThermoFisher Scientific, Cat# A-11057, RRID:AB_142581, 1:200), AlexaFluor 647 F(ab’) 2 fragment donkey anti-rat IgG (Jackson ImmunoResearch, Cat# 712-606-153, RRID:AB_2340696, 1:200), and AlexaFluor 647 donkey anti-rabbit IgG (Abcam, Cat# ab150075, RRID:AB_2752244, 1:400).

Techniques: Staining, Isolation