Mouse/Rat Neuropilin-1 Alexa Fluor® 700-conjugated Antibody

Detection of Neuropilin‑1 in bEnd.3 Mouse Cell Line by Flow Cytometry. bEnd.3 mouse endothelioma cell line was stained with Goat Anti-Mouse/Rat Neuropilin-1 Alexa Fluor® 700-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB566N, filled histogram) or isotype control antibody (Catalog # IC108N, open histogram). View our protocol for Staining Membrane-associated Proteins.

Detection of Neuropilin‑1 in PC‑12 Rat Cell Line by Flow Cytometry. PC-12 rat adrenal pheochromocytoma cell line was stained with Goat Anti-Mouse/Rat Neuropilin-1 Alexa Fluor® 700-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB566N, filled histogram) or isotype control antibody (Catalog # IC108N, open histogram). View our protocol for Staining Membrane-associated Proteins.

Detection of Mouse Neuropilin-1 by Flow Cytometry pTregs show a higher susceptibility to convert into exFoxp3 cells after transfer into dry eye hosts. pTregs (CD4+CD25+Nrp-1−) and tTregs (CD4+CD25+Nrp-1+) from transgenic dry eye disease (DED) mice (Foxp3-GFP×R26-RFP) were adoptively transferred into control and DED transplant recipients. (A and B) Draining lymph nodes were harvested at day 14 post-transplantation. Single cell suspensions were analyzed by flow cytometry. Frequencies of exFoxp3 cells (GFP−RFP+) from control and DED hosts that received (A) pTregs (59.02 ± 0.92% vs. 78.26 ± 0.69%, n = 4, ***p < 0.0001 unpaired two-tailed Student’s t test) or (B) tTregs was assessed. (C) Representative flow cytometry plots showing frequencies of IL-17 and IFN gamma -expressing exFoxp3 cells in draining lymph nodes from control and dry eye hosts. (D) Nrp1+ tTregs or Nrp1− pTregs from hosts with accepted allografts were adoptively transferred to dry eye hosts, and graft survival was assessed for 8 weeks (n = 10, χ2 = 0.6087, p = 0.4353). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29728574), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Neuropilin-1 by Flow Cytometry Dry eye disease promotes loss of Foxp3 expression by Tregs following corneal transplantation. Foxp3-GFP-CreXR26-RFP transgenic mice were used to trace Foxp3 expression after transplantation. Healthy corneal grafts were transplanted onto transgenic naïve control hosts or dry eye (DED) hosts. Draining lymph nodes were harvested 2 weeks post-transplantation. Single cell suspensions were analyzed by flow cytometry. (A) Representative flow cytometry plots and (B) statistical analysis of flow cytometry data showing frequencies of exFoxp3 cells in Foxp3-GFPXR26-RFP control and DED hosts (n = 6/group, ***p < 0.001). (C) Representative flow cytometry plots and (D, E) statistical analyses of flow cytometry data showing frequencies of exFoxp3 cells that express IFN gamma (***p < 0.001) and IL-17 (**p < 0.01) in Foxp3-GFPXR26-RFP control and DED hosts (n = 6/group). Data is presented as mean ± SEM (error bar). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29728574), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Neuropilin-1 by Flow Cytometry Dry eye disease reduces graft survival and Treg function after allograft survival. Healthy corneal grafts were transplanted on normal control hosts or dry eye (DED) hosts. (A) Corneal graft opacity (***p < 0.001) and (B) graft survival were assessed for 8 weeks (n = 10/group, **p < 0.01). Draining lymph nodes were harvested 2 weeks post-transplantation. Single cell suspensions were analyzed by flow cytometry. (C) Frequencies of CD4+Foxp3+ Tregs from control vs. DED hosts was determined using flow cytometry (n = 5 and ***p < 0.001) (D) Treg suppression assay (n = 6) was performed using CD4+CD25− responders from rejected control hosts, and CD4+CD25+Tregs from control and dry eye hosts (**p < 0.01). Data is presented as mean ± SEM (error bar). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29728574), licensed under a CC-BY license. Not internally tested by R&D Systems.