vegf c elisa  (Novus Biologicals)

Journal: Stem Cell Research & Therapy

Article Title: Thrombopoietin mimetic stimulates bone marrow vascular and stromal niches to mitigate acute radiation syndrome

doi: 10.1186/s13287-024-03734-z

Figure Lengend Snippet: TPOm systemically reduces bone marrow vascular dilatation and vascular leakage and promotes production of VEGF-A and VEGF-C in irradiated mice. A Representative immunofluorescent images of femurs stained with DAPI (blue), VEGFR3 (green), and CD31/CD144 (red) on days 4 and 10 after irradiation in vehicle and TPOm-treated mice. Non-irradiated mice were represented as naïve for reference. Scale bar is 10 μm. B VEGFR3 + vessel area in the bone marrow on days 4 and 10 after irradiation, quantitated by using Volocity software (n = 3/group). C Representative immunofluorescent images of sternum stained with DAPI (blue), Sca-1 (green), and TIE2 (red) on days 2 and 4 after irradiation in vehicle and TPOm-treated mice from 2 independent experiments. Non-irradiated mice were represented as naïve for reference. D Quantification of Sca-1 + cells per 100 × field in sternal bone marrow quantitated by using Volocity software (n = 3/group). E IVIS images of 7.2 Gy (X-rays) TBI mice imaged 5 days after irradiation with AngioSense750 EX i.v. injection performed on day 3 after irradiation. F Quantification of total radiant efficiency (n = 3/group). G , H ELISA of VEGF-A in ( G ) serum and in ( H ) BM after 7.0 Gy ( 137 Cs) TBI. I , J ELISA of VEGF-C in ( I ) serum and in ( J ) BM after 7.0 Gy ( 137 Cs) TBI. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01 vehicle vs . TPOm-treated assessed by unpaired Student’s t -test with post hoc Holm-Sidak method for multiple comparisons

Article Snippet: , VEGF-C ELISA , Novus Biologicals , Cat# NBP2-78893, RRID: AB_3083672.

Techniques: Irradiation, Staining, Software, Injection, Enzyme-linked Immunosorbent Assay

Journal: Stem Cell Research & Therapy

Article Title: Thrombopoietin mimetic stimulates bone marrow vascular and stromal niches to mitigate acute radiation syndrome

doi: 10.1186/s13287-024-03734-z

Figure Lengend Snippet:

Article Snippet: , VEGF-C ELISA , Novus Biologicals , Cat# NBP2-78893, RRID: AB_3083672.

Techniques: Cell Analysis, Recombinant, Electron Microscopy, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, Software, Imaging

Journal: bioRxiv

Article Title: Langerhans cells regulate immunity in adulthood by regulating postnatal dermal lymphatic development

doi: 10.1101/2024.07.12.603312

Figure Lengend Snippet: ( A-B ) 3 week old mice of indicated genotypes were treated intradermally in ear skin with indicated blocking antibodies for 2-3 ( A ) and with indicated cytokines for 2 ( B ) weeks before ear LECs were quantitated. Representative plots (left), absolute and relative LEC numbers (right). ( C, D ) Murine dermal LECs were cultured with or without LCs and with anti-PIGF, anti-VEGFR3, or anti-PIGF+anti-VEGFR3 before LECs were quantitated. Relative LEC numbers ( C ) and percentage that are DAPI+ ( D ). ( E-F ) mRNA expression of Pgf ( E ) and Vegfc ( F ) by sorted LCs, T cells, and keratinocytes (KCs) from 4 and 8 week old WT mice. Positive control (ctrl) was murine placenta. ( G ) LCs from 4 and 8 week old WT mice were cultured for 2 days and supernatant was assessed for VEGF-C by ELISA. ( H ) VEGF-C cell binding. LCs and other cells in the medium after a 48-hour crawl-out from 4 and 8 week old mouse epidermal sheets were incubated with his-tagged VEGF-C which was detected with anti-his tag. Representative histograms (left) and percentage of indicated cells that bound his-VEGF-C (right). ( I ) Ear skin LECs in 8 week old WT and CCR7 KO mice were quantitated. Percentage (left) and number of LECs (right). Each symbol represents one mouse. Bars represent mean and error bars s.e.m. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 using two-tailed paired ( A , left) or unpaired Student’s t-test ( A , right, B-I ). Mice per condition: n= 4 ( A ), 4-6 ( B ), 4-5 ( C-D ), 3-5 ( E-F ), 5 ( G ), 5-6 ( H ), 6 ( I ) and data are from 2 of multiple similar experiments ( H ) and 3 ( A,C-F ), 4 ( G, I ) and 5 ( B ) independent experiments.

Article Snippet: Culture supernatants were analyzed for VEGF-C using a VEGF-C ELISA kit (CUSABIO, #CSB-E07361M, Houston, TX).

Techniques: Blocking Assay, Cell Culture, Expressing, Positive Control, Enzyme-linked Immunosorbent Assay, Binding Assay, Incubation, Two Tailed Test

Journal: Nature Neuroscience

Article Title: Piezo1 regulates meningeal lymphatic vessel drainage and alleviates excessive CSF accumulation

doi: 10.1038/s41593-024-01604-8

Figure Lengend Snippet: a , Increased phosphorylation of CDH5 (Try 658), VEGFR2 (Tyr 1054/Tyr 1059) and VEGFR3 (Tyr 1230/Tyr 1231) in cultured primary human LECs in vitro treated with Yoda1 (0.5 µM, final). b , CDH5 immunofluorescence stains showing increased junctional gaps (arrowheads) in 2D LEC in vitro cultures after Yoda1 treatment at 0.5, 1 or 2 µM for 8 h; scale bars, 20 µm. c , Piezo1-dependent increase in drainage efficiency of engineered lymphatics in vitro. Primary LECs were transfected with control (scrambled) or Piezo1 short interfering RNA (siRNA) for 24 h, used to build lymphatics in 3D PDMS chips and treated with vehicle or Yoda1 (1 µM, final) to evaluate drainage capability. Detailed images for the engineered lymphatics are shown in Extended Data Fig. ; siCTR and vehicle, n = 7 independent experiments; siCTR and Yoda1, n = 8 independent experiments; siPiezo1 and vehicle, n = 8 independent experiments; siPiezo1 and Yoda1, n = 5 independent experiments. Data were analyzed by one-way ANOVA ( P = 0.0097) followed by a Bonferroni multiple comparison test. d , Lymphatic vessel contractility test. Surgically excised axillary collecting lymphatics were treated with vehicle, Yoda1 or l -NAME + Yoda1 at the indicated intraluminal pressures, and the percent vessel tone was measured. Additional functionality values are shown in Extended Data Fig. ; n = 6 independent samples. Data were analyzed by two-way repeated measures ANOVA ( P = 0.003) between treatments followed by Tukey’s multiple comparison test; P < 0.0001 between control and Yoda1; P = 0.0004 between control and l -NAME + Yoda1; P < 0.0001 between Yoda1 and l -NAME + Yoda1. e , Inhibition of Piezo1 downstream effectors suppresses Yoda1-induced promotion of brain tracer drainage. Fluorescent tracer (OVA-Green) was premixed with vehicle, Yoda1 (50 µM, final) or Yoda1 (50 µM, final) + inhibitors, such as l -NAME (eNOS), capivasertib (Cap; AKT), axitinib (Axi; VEGFR1–VEGFR3), SAR131675 (SAR; VEGFR3) or cabozantinib malate (Cab; VEGFR2; 258 µg ml –1 final concentration of each inhibitor), and i.c.m. injected into Prox1 -tdTomato mice (15 weeks old). After 45 min, LNs were collected and imaged; scale bars, 500 µm. f , g , Quantification of relative tracer intensity in cervical ( f ) and mandibular ( g ) LNs ( n = 7 mice per group). One data point represents the sum of the fluorescence intensity of LNs of both sides of a single mouse. Data were analyzed by one-way ANOVA ( P < 0.0001) followed by Tukey’s multiple comparison test. h , i , Protein expression levels of VEGF-C ( h ) and VEGF-D ( i ) were measured in the whole brains of mice that were treated with vehicle or Yoda1 (213 µg per kg (body weight) two times at 18 h and 90 min before tissue collection) or in the whole brains of control or Piezo1 TG_LEC mice that were i.p. injected with tamoxifen (50 mg per kg (body weight) twice, 3 days apart) at the age of 8 weeks. Pz1 TG , Piezo1 TG_LEC mice. Each data point represents one mouse ( n = 5 mice per group). Data were analyzed by two-tailed t -test and are presented as mean values ± s.e.m.

Article Snippet: Concentrations of VEGF-C and VEGF-D in the brain extracts were quantified using a mouse VEGF-C ELISA kit and a mouse VEGF-D ELISA kit (CUSABIO, CSB-E07361m and CSB-E07357m, respectively).

Techniques: Cell Culture, In Vitro, Immunofluorescence, Transfection, Small Interfering RNA, Comparison, Inhibition, Concentration Assay, Injection, Fluorescence, Expressing, Two Tailed Test