mouse vascular endothelial cell growth factor c  (Cusabio)

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: bioRxiv

Article Title: Immunoregulatory subtype of dermal lymphatic endothelial cells at capillary terminals drives lymphatic malformations

doi: 10.1101/2022.05.22.492950

Figure Lengend Snippet: ( A-E ) Quantification of the CD45 + area ( A , B ) and F4/80 + area ( C - E ) in the ear skin showing increase in Pik3ca H1047R ;Vegfr3-CreER T2 ( A, C, D ) but not in Pik3ca H1047R ;Vegfr1-CreER T2 ( B, C, E ) mice. Data represent mean (CD45: n =4-6 images from n =3-5 mice per genotype; F4/80: n =3-7 images from n =2-3 mice per genotype) ± s.e.m. Representative binary images are shown below the graphs. ( F, G ) Flow cytometry analysis of innate ( F ) and adaptive ( G ) immune cells in the ear skin of 4-OHT-treated 5-week-old and 10-week-old Pik3ca H1047R ; Vegfr3-CreER T2 mice and littermate controls. Mac, macrophage; Mono, monocyte; DC, dendritic cell; Neu, neutrophil; NK, natural killer cell. Data represent relative cell frequency (of live cells) relative to the control ( n =3-9 mice) ± s.e.m. ( H ) Multiplex ELISA analysis of pro-inflammatory cytokines and chemokines associated with recruitment and/or activation of myeloid cells in whole ear skin lysates from Pik3ca H1047R ;Vegfr3-CreER T2 mice and littermate controls. Data represent mean protein levels relative to control ( n =6-11 mice) ± s.e.m. ( I ) qRT-PCR analysis (left) and ELISA analysis (right) of Vegfc /VEGF-C levels in the ear skin lysates of 4-OHT-treated 5-week-old Pik3ca H1047R ;Vegfr3-CreER T2 and littermate control mice. qRT-PCR data represent mean relative expression (normalized to Hprt ; n =7-10 mice) ± s.e.m. Transcript and protein levels are presented relative to controls. ( J ) qRT-PCR analysis of Vegfc in CD45 + Cd11b + F4/80 + macrophages and CD45 + Cd11b + F4/80 + Ly6C + monocytes, FACS-sorted from the ear skin of 4-OHT-treated 5-week-old Pik3ca H1047R ;Vegfr3-CreER T2 and littermate control mice. qRT-PCR data represent mean relative expression (normalized to Hprt ; n =3-5 mice) ± s.e.m. Transcript and protein levels are presented relative to macrophages/monocytes in control mice. p -value in (A, B, D-I) obtained using Two-tailed unpaired Student’s t-test. Scale bar: 100 μm (A-E).

Article Snippet: Mouse Vascular Endothelial Cell Growth Factor C (VEGF-C) ELISA Kit from CUSABIO (CSB-E07361m) was used for detection of VEGF-C protein concentration.

Techniques: Flow Cytometry, Control, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Activation Assay, Quantitative RT-PCR, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Immunoregulatory subtype of dermal lymphatic endothelial cells at capillary terminals drives lymphatic malformations

doi: 10.1101/2022.05.22.492950

Figure Lengend Snippet: ( A ) Flow cytometry analysis of the number of CD45 + CD11b + F4/80 + macrophages in the ear skin of 4-OHT-treated 5-week-old Pik3ca H1047R ;Vegfr3-CreER T2 ( n =5) and control ( n =3) mice. Data represent mean cell number per gram tissue ± s.e.m. p -value, Mann-Whitney U Test. ( B ) Flow cytometry analysis of the frequency of CD45 + CD11b + F4/80 + antigen-presenting myeloid cells in the ear skin of 4-OHT-treated 5-week-old Pik3ca H1047R ;Vegfr1-CreER T2 ( n =6) and control ( n =5) mice. Data represent relative cell frequency (of live cells) relative to the control ± s.e.m. p -value obtained using Two-tailed unpaired Student’s t-test. ( C ) Flow cytometry analysis of innate (left) and adaptive (right) immune cells in the ear skin of 4-OHT-treated 10-week-old Pik3ca H1047R ; Vegfr1-CreER T2 mice and littermate controls. Mac, macrophage; Mono, monocyte; DC, dendritic cell; Neu, neutrophil; NK, natural killer cell. Data represent relative cell frequency (of live cells) relative to the control ( n =5-8 mice for innate panel, n=7-9 mice for adaptive panel) ± s.e.m. p -value obtained using Two-tailed unpaired Student’s t-test. ( D, E ) Multiplex ELISA analysis of pro-inflammatory cytokines and chemokines associated with recruitment and/or activation of myeloid cells or T-cells and B-cells in whole ear skin lysates from Pik3ca H1047R ;Vegfr1-CreER T2 ( D ) and Pik3ca H1047R ;Vegfr3-CreER T2 ( E ) mice, and respective littermate controls. ( F ) Similar analysis of TNFα and INFγ in blood serum of Pik3ca H1047R ;Vegfr3-CreER T2 mice. Data in ( D-E ) represent mean protein levels ( n =3-9 mice) ± s.e.m.

Article Snippet: Mouse Vascular Endothelial Cell Growth Factor C (VEGF-C) ELISA Kit from CUSABIO (CSB-E07361m) was used for detection of VEGF-C protein concentration.

Techniques: Flow Cytometry, Control, MANN-WHITNEY, Two Tailed Test, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Activation Assay