Journal: Fluids and Barriers of the CNS

Article Title: Compartmentalized VEGF receptor expression in hypothalamic tanycytes reveals a novel non-endothelial axis of VEGF signaling

doi: 10.1186/s12987-026-00774-w

Figure Lengend Snippet: Single cell RNA sequencing of VEGFR family in tanycytes. ( A ) On the left: Schematic of the ME representing its privileged localisation in the hypothalamus. On the right: Correlation between tanycyte subtypes and their corresponding hypothalamic nuclei. Tanycytes are color coded according to the UMAP in B. ( B ) UMAP showing clusters representative of four main tanycytic subpopulations generated by the reanalysis of scRNA-seq ME dataset from [18]. ( C ) Feature plots showing VEGFRs (Kdr, Flt1 and Flt4), ligands (Vegfa, Vegfb and Vegfc) and co-receptors (Nrp1 and Nrp2) in tanycytic subclusters. ( D ) Dot plot showing the expression of the different members of the family across the tanycytic subclusters

Article Snippet: VEGFc , Mm00437310_m1.

Techniques: Single Cell, RNA Sequencing, Generated, Expressing

Journal: Fluids and Barriers of the CNS

Article Title: Compartmentalized VEGF receptor expression in hypothalamic tanycytes reveals a novel non-endothelial axis of VEGF signaling

doi: 10.1186/s12987-026-00774-w

Figure Lengend Snippet: Quantification of Vegfr1, Vegfr2 and Vegfa during postnatal development. ( A ) qPCR analysis of Vegfr1, Vegfr2, and Vegfr3 mRNA expression levels in tanycytes isolated from male mice aged 3, 6, 9, 12, and 18 months. Notably, Vegfr2 expression shows a progressive decline with age, while Vegfr1 and Vegfr3 remain stable. ( B ) qPCR analysis of VEGFR ligands. Vegfa and Vegfc remain relatively stable across age, while Vegfb shows a modest upregulation in older mice. ( C ) Expression of VEGF co-receptors Nrp1 and Nrp2 in sorted tanycytes. While Nrp1 remains unchanged, Nrp2 expression declines with age, paralleling the pattern of Vegfr2. qPCR data are normalized to housekeeping genes 18S and Actb, and presented as mean ± SEM; n = 5–6 mice per time point. Statistical analysis was performed using one-way ANOVA with Fisher’s LSD post hoc test. p < 0.05 was considered significant

Article Snippet: VEGFc , Mm00437310_m1.

Techniques: Expressing, Isolation

Journal: Experimental Biology and Medicine

Article Title: Engineering ADSCs by manipulating YAP for lymphedema treatment in a mouse tail model

doi: 10.3389/ebm.2024.10295

Figure Lengend Snippet: VEGFC successfully induced lymphatic endothelial transdifferentiation of ADSCs. (A–C) PCR tests showed the upregulation of VEGFR-3, Prox-1, Lyve-1 during the lymphatic endothelial transdifferentiation of ADSCs. (D) VEGFR-3 as a typical LEC marker was detected by immunofluorescence staining after VEGFC-induction at the indicated times. Scale bar = 50 μm. (E) Immunofluorescence intensity analysis of VEGFR-3. (F, H) Western blot showed the increased VEGFR-3 expression during the lymphatic endothelial transdifferentiation of ADSCs. (G) ADSCs were seeded on Matrigel after 7-day induction, and tube formation was evaluated at 12 h postseeding. VEGFC group generated tube-like structure while control group did not exhibit tubes. Scale bar = 100 μm. Bars: means ± standard deviation. n = 3 in each group, ns: no significant, ** P < 0.01, *** P < 0.001.

Article Snippet: For the lymphatic transdifferentiation of ADSCs, VEGFC (100 ng/mL, HY-P77864, MedChem Express, Monmouth Junction, NJ, United States) was used for 7 days.

Techniques: Marker, Immunofluorescence, Staining, Western Blot, Expressing, Generated, Control, Standard Deviation

Journal: Experimental Biology and Medicine

Article Title: Engineering ADSCs by manipulating YAP for lymphedema treatment in a mouse tail model

doi: 10.3389/ebm.2024.10295

Figure Lengend Snippet: Effect of lymphatic endothelial transdifferentiation and verteporfin on the expression of YAP. (A) Immunostaining of YAP in the control group and VEGFC group. Scale bar = 50 μm. (B) Immunofluorescence intensity analysis of YAP. (C–E) Western blot showed the decreased nuclear YAP expression and increased cytosolic YAP expression in ADSCs after lymphatic endothelial transdifferentiation. (F) PCR test showed that verteporfin suppressed the expression of YAP in ADSCs at the concentration of 20 μM. (G , H) Western blot showed the continuously inhibitory effect of verteporfin on YAP expression in ADSCs. Bars: means ± standard deviation. n = 3 in each group; ns, no significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For the lymphatic transdifferentiation of ADSCs, VEGFC (100 ng/mL, HY-P77864, MedChem Express, Monmouth Junction, NJ, United States) was used for 7 days.

Techniques: Expressing, Immunostaining, Control, Immunofluorescence, Western Blot, Concentration Assay, Standard Deviation

Journal: Experimental Biology and Medicine

Article Title: Engineering ADSCs by manipulating YAP for lymphedema treatment in a mouse tail model

doi: 10.3389/ebm.2024.10295

Figure Lengend Snippet: The downregulation of YAP enhanced the lymphatic endothelial transdifferentiation of ADSCs in vitro . 20 μM verteporfin preconditioning for 48 h downregulated the expression of YAP in ADSCs. Under this inhibitory effect, higher expression levels of VEGFR-3 can be detected after differentiation of ADSCs, and larger density of tube formation can be observed. (A) Lymphatic endothelial transdifferentiation of ADSCs was conducted after 20 μM-verteporfin preconditioning for 48 h, and immunofluorescence staining indicated higher level of VEGFR-3. Scale bar = 50 μm. (B) Immunofluorescence intensity analysis of VEGFR-3. (C, D) Higher expression level of VEGFR-3 was confirmed by western blot. (E, F) The tube formation assay showed that VEGFC (+) verteporfin (+) group generated more tube-like structure than VEGFC (+) verteporfin (−) group. And quantification of master segments was analysed. Scale bar = 200 μm. Bars: means ± standard deviation. n = 3 in each group, ns: no significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For the lymphatic transdifferentiation of ADSCs, VEGFC (100 ng/mL, HY-P77864, MedChem Express, Monmouth Junction, NJ, United States) was used for 7 days.

Techniques: In Vitro, Expressing, Immunofluorescence, Staining, Western Blot, Tube Formation Assay, Generated, Standard Deviation

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Therapeutic Lymphangiogenesis With Implantation of Adipose‐Derived Regenerative Cells

doi: 10.1161/JAHA.112.000877

Figure Lengend Snippet: The ability of ADRCs for lymphangiogenesis in vitro (functional assay and differentiation assay). A, Enzyme‐linked immunosorbent assay (ELISA) revealed that the concentration of VEGF‐C was upregulated in ADRC‐CM compared to control. B, rhVEGF‐C induced LEC migration in a dose‐dependent manner, and ADRC‐CM also induced LEC migration. HPF indicates high‐powered field. * P <0.05 vs nontreated control cells. C, Proliferation assay also revealed that ADRC‐CM promoted LEC proliferation, but VEGF‐C did not. * P <0.05, ** P <0.01 vs control. D, Western blot analysis revealed that ADRC‐CM, as well as VEGF‐C, had an ability to phosphorylate Erk and eNOS in LECs. Western blot analysis showed that phosphorylated Erk (p‐Erk) and phosphorylated eNOS (p‐eNOS) expression was greater in the VEGF‐C– or ADRC‐CM–treated LECs than in control nontreated LECs. E, Cultured with growth factors, the expression of podoplanin mRNA was greater than that of control. Immunocytochemistry also revealed that cultured ADRCs were stained with LEC marker.

Article Snippet: Concentrations of VEGF‐C proteins in the media were determined by a mouse VEGF‐C enzyme‐linked immunosorbent assay kit (Cusabio Biotech Co, Ltd) according to the manufacturer's instructions.

Techniques: In Vitro, Functional Assay, Differentiation Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Control, Migration, Proliferation Assay, Western Blot, Expressing, Cell Culture, Immunocytochemistry, Staining, Marker

Journal: REPRODUCTION

Article Title: Possible involvement of IFNT in lymphangiogenesis in the corpus luteum during the maternal recognition period in the cow

doi: 10.1530/rep-11-0157

Figure Lengend Snippet: Figure 6 Effects of IFNTor IFNA on ISG15, VEGFC, and VEGFR3 mRNA expression in LyECs and LECs. IFNT stimulated mRNA expression of ISG15 and VEGFC but not VEGFR3 in LyECs (A–C). IFNT also stimulated ISG15 and VEGFC mRNA expression in LECs (D and E). In contrast, IFNA stimulated ISG15 mRNA expression (F) but tended to decrease VEGFC mRNA expression in LyECs (G). IFNA had no effect on VEGFR3 mRNA expression (H). In LECs, IFNA stimulated ISG15 mRNA expression but not VEGFC mRNA expression (I and J). White bars indicate the control group (no treatment), while black bars indicate 0.6–60 IU/ml IFNT-treated or IFNA-treated groups. All values are shown as meanGS.E.M. (nZ4/group). *Significant differences (P!0.05) and #tendency to difference (P!0.1) as determined by ANOVA followed by Fisher’s multiple comparison test.

Article Snippet: Inc. (Otsu, Japan); 4% Block Ace Powder was purchased from DS Pharma Biomedical (Osaka, Japan); anti-mouse-LYVE1 rabbit polyclonal antibody was purchased from Abcam (Cambridge, UK); BD Matrigel basement membrane was purchased from BD Biosciences (Bedford, MA, USA); recombinant human VEGFC was purchased from ProSci (Poway, CA, USA); protease inhibitor cocktail was purchased from Roche; 70 mm filter (Cell Straner, REF 352350) was purchased from BD Falcon (Franklin Lakes, NJ, USA); PVDF membranes were purchased from Bio-Rad Laboratories; lymphoprep was purchased from AxisShield (Oslo, Norway); HRP-conjugated anti-rabbit IgG antibodies were purchased from GE Healthcare Ltd (Chalfont St Giles, UK); HRP-conjugated anti-mouse IgG antibodies were purchased from Rockland Immunochemicals, Inc. (Gilbertsville, PA, USA); anti-human VWF rabbit polyclonal antibody (clone A0082) was purchased from Dako Denmark A/S (Glostrup, Denmark); biotinylated goat anti-rabbit BA-1000 IgG and avidin–biotin reagent (PK-6100, Vectastain ABC kit) were purchased from Vector Laboratories, Inc. (Burlingame, CA, USA); recombinant bovine IFN was produced by Escherichia coli.

Techniques: Expressing, Control, Comparison

Journal: REPRODUCTION

Article Title: Possible involvement of IFNT in lymphangiogenesis in the corpus luteum during the maternal recognition period in the cow

doi: 10.1530/rep-11-0157

Figure Lengend Snippet: Figure 7 Effects of IFNT on proliferation of LyECs and capillary-like tube formation. IFNT stimulated the proliferation of LyECs (A) and promoted the capillary-like tube formation of LyECs as well as 100 ng/ml VEGFC (B). Panels C, D and E show typical images of capillary-like tube formation in matrigel assays on LyECs. All values are shown as meanGS.E.M. (nZ4/group). The scale bars rep- resent 500 mm. *, **Significant differences (P!0.05 or P!0.01) and #tendency to difference (P!0.1) as determined by ANOVA followed by Fisher’s multiple comparison test.

Article Snippet: Inc. (Otsu, Japan); 4% Block Ace Powder was purchased from DS Pharma Biomedical (Osaka, Japan); anti-mouse-LYVE1 rabbit polyclonal antibody was purchased from Abcam (Cambridge, UK); BD Matrigel basement membrane was purchased from BD Biosciences (Bedford, MA, USA); recombinant human VEGFC was purchased from ProSci (Poway, CA, USA); protease inhibitor cocktail was purchased from Roche; 70 mm filter (Cell Straner, REF 352350) was purchased from BD Falcon (Franklin Lakes, NJ, USA); PVDF membranes were purchased from Bio-Rad Laboratories; lymphoprep was purchased from AxisShield (Oslo, Norway); HRP-conjugated anti-rabbit IgG antibodies were purchased from GE Healthcare Ltd (Chalfont St Giles, UK); HRP-conjugated anti-mouse IgG antibodies were purchased from Rockland Immunochemicals, Inc. (Gilbertsville, PA, USA); anti-human VWF rabbit polyclonal antibody (clone A0082) was purchased from Dako Denmark A/S (Glostrup, Denmark); biotinylated goat anti-rabbit BA-1000 IgG and avidin–biotin reagent (PK-6100, Vectastain ABC kit) were purchased from Vector Laboratories, Inc. (Burlingame, CA, USA); recombinant bovine IFN was produced by Escherichia coli.

Techniques: Comparison