Journal: Vascular Biology

Article Title: Penile cavernous sinusoids are Prox1-positive hybrid vessels

doi: 10.1530/VB-23-0014

Figure Lengend Snippet: Characterization of the murine penile vasculature. (A) Schematic presentation of the murine penile anatomy. Blood vessels (pink), penile cavernous sinusoids (pc-Ss) (red), lymphatic vessels (green), baculum (penis bone, gray), and ureter (brown) are displayed. CCg, corpus cavernosum glandis; CCug, corpus cavernosum urethrae glandis; CCu, corpus cavernosum urethrae; CC, corpus cavernosum. (B, C) Vascular blood perfusion (visualized by lectin perfusion) and expression of Prox1-GFP in the penile glans and body. A magnified view of the boxed region in (B) is shown in (C). Red arrowheads point to blood-perfused Prox1-GFP -positive sinusoids, and blood vessels (red arrows) and lymphatic vessels (green arrowheads) are indicated. (D–F) Imaris rendering from two-photon images of cavernous tissue from 12-month-old R26‑mTmG;Vegfr3-CreER T2 mice, treated with 1 mg of tamoxifen on three consecutive days at 11 months of age, displayed in colored depth coding and showing different angles of a coronal section of CC (arrow) (D). Note colocalization of GFP ( Vegfr3 expression) with intravenously injected lectin (purple) in CC and CCu but not in lymphatic vessels (LV) (E). (F) CCu (arrowhead) and CC (arrow) are displayed. Scale bars, 50 μm (C), 80 μm (D), 100 μm (F (right panel)), 200 μm (B, E, F (left panel)).

Article Snippet: Recombinant mouse VEGFR3 chimera protein (R&D Systems, 743-R3-100) was used as quantitative standard.

Techniques: Expressing, Injection

Journal: Vascular Biology

Article Title: Penile cavernous sinusoids are Prox1-positive hybrid vessels

doi: 10.1530/VB-23-0014

Figure Lengend Snippet: Endothelial cell marker expression in penile cavernous sinusoids (pc-Ss). The analyzed markers in ECs of pc-Ss in comparison to previously reported expression in hybrid vessels: Schlemm’s canal (SC) ( 2 , 3 , 4 , 56 , 57 , 58 , 59 , 60 , 61 ), ascending vasa recta (AVR) ( 5 , 50 , 62 , 63 ), and remodeled spiral artery (rSA) ( 6 , 64 , 65 , 66 , 67 ).

Article Snippet: Recombinant mouse VEGFR3 chimera protein (R&D Systems, 743-R3-100) was used as quantitative standard.

Techniques: Marker, Expressing, Comparison

Journal: Sarcoma

Article Title: Axitinib Has Antiangiogenic and Antitumorigenic Activity in Myxoid Liposarcoma

doi: 10.1155/2016/3484673

Figure Lengend Snippet: eIF4E promotes angiogenic properties. MLS 402 and MLS 1765 cells were treated with IC 50 4EGI-1 (MLS 402, 8.2 μ M; MLS 1765, 4.8 μ M) or vehicle control (VC) overnight, and then RT-qPCR was performed to measure the expression of VEGF ligands and receptors. The assay was performed three times. The data shown are from three independent experiments presented as mean + SEM relative to the housekeeping gene H6PD (a). HUVECs were suspended in Matrigel, and then conditioned medium from MLS 402 cells pretreated with IC 50 4EGI-1 or vehicle control overnight was applied. Images were acquired at hourly intervals, and the figure displays representative images taken at 8 h. Tube length was measured using ImageJ (version 1.47d) (b). CM, conditioned medium; IB, immunoblotting; LO, Lipofectamine-only treated cells; Ut, untreated MLS cells. MLS 402 and MLS 1765 cells were plated overnight and then treated with 20 μ g/mL VEGFR1 ligand trap (LT), VEGFR3 ligand trap, both ligand traps, or vehicle control for 3 days. Then an MTS dye uptake assay was performed to measure cell viability, and the effect on cell proliferation was calculated (c). Technical duplicates and biological triplicates were tested. A paired, two-tailed t -test was performed. The data are presented as mean + SEM. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Myxoid liposarcoma cells were seeded at a density of 1,000–2,000 cells per well on 96-well plates and incubated at 37°C in 5% CO 2 for 18 h. Then ligand traps were added at 2.0 μ g/100 μ L: recombinant human VEGFR1 (FLT-1)-Fc chimera (R&D Systems), recombinant mouse VEGFR3 (FLT-4)-Fc chimera (R&D Systems), both of these ligand traps, or vehicle control.

Techniques: Control, Quantitative RT-PCR, Expressing, Western Blot, Two Tailed Test

Journal: Sarcoma

Article Title: Axitinib Has Antiangiogenic and Antitumorigenic Activity in Myxoid Liposarcoma

doi: 10.1155/2016/3484673

Figure Lengend Snippet: Characterization of axitinib. MLS 402 and MLS 1765 cells were treated with IC 50 axitinib (MLS 402, 1.2 μ M; MLS 1765, 3.2 μ M), imatinib (10 μ M), or vehicle control for 2 h, lyzed, and analyzed for anolytes targeting the phosphorylation site of PDGFR α , PDGFR β , and c-Kit. The experiment was performed 3 times, and the data are presented as mean + SEM. t-AKT, total AKT (a). MLS 1765 cells were treated overnight with IC 50 axitinib or vehicle control and immunoprecipitated for total VEGFR3 (t-VEGFR3) and phospho-VEGFR3 (p-VEGFR3). Densitometry of total VEGFR3 expression and phospho-VEGFR3 expression following axitinib treatment (b). MLS 402 and MLS 1765 cells were treated with IC 50 axitinib or vehicle control for 2 h, and then the cells were lyzed and analyzed for phospho-AKT and phospho-ERK1/2 by Bio-Plex. Three technical and biological replicates were tested. The data are presented as the mean + SEM (c). MLS 402 and MLS 1765 cells were treated overnight with IC 50 axitinib or vehicle control, and then cells were analyzed by RT-qPCR for the expression of VEGFR1, VEGFR3, VEGFA, and VEGFB. The experiment was performed three times, and the data are presented as the mean + SEM (d). HUVECs were suspended in Matrigel and then were treated with conditioned medium from cells that had been pretreated with IC 50 axitinib or vehicle control. Images were acquired at hourly intervals, and the figure displays representative images taken at 8 h. Tube lengths were measured using ImageJ (version 1.47d). The experiment was performed three times, and the data are presented as the mean + SEM (right) (e). Ax, axitinib; CM, conditioned medium; IB, immunoblotting; Im, imatinib; IP, immunoprecipitation; VC, vehicle control; LO, Lipofectamine-only treated cells; Ut, untreated MLS cells. A paired, two-tailed t -test was performed. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Myxoid liposarcoma cells were seeded at a density of 1,000–2,000 cells per well on 96-well plates and incubated at 37°C in 5% CO 2 for 18 h. Then ligand traps were added at 2.0 μ g/100 μ L: recombinant human VEGFR1 (FLT-1)-Fc chimera (R&D Systems), recombinant mouse VEGFR3 (FLT-4)-Fc chimera (R&D Systems), both of these ligand traps, or vehicle control.

Techniques: Control, Phospho-proteomics, Immunoprecipitation, Expressing, Quantitative RT-PCR, Western Blot, Two Tailed Test