Journal: Molecular medicine reports

Article Title: Intraperitoneal injection of thalidomide alleviates early osteoarthritis development by suppressing vascular endothelial growth factor expression in mice.

doi: 10.3892/mmr.2018.8980

Figure Lengend Snippet: Figure 2. mRNA expression levels of VEGF and MMP‑13 in the knee articular cartilage of mice among the Sham, Dmm and Dmm+Th groups (n=4 in each group). (A) Relative mRNA expression levels of VEGF in the medial articular cartilage. (B) Relative mRNA expression levels of MMP‑13 in the medial articular cartilage. The values are presented as the mean ± standard deviation. *P<0.05 compared with the Sham group; #P<0.05 compared with the Dmm group. Dmm, destabilization of the medial meniscus; MMP‑13, matrix metalloproteinase‑13; Th, thalidomide; VEGF, vascular endothelial growth factor.

Article Snippet: An ELISA kit of VEGF (E-EL-M1292c) was purchased from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Expressing, Standard Deviation

Journal: Molecular medicine reports

Article Title: Intraperitoneal injection of thalidomide alleviates early osteoarthritis development by suppressing vascular endothelial growth factor expression in mice.

doi: 10.3892/mmr.2018.8980

Figure Lengend Snippet: Figure 3. Immunohistochemical analysis of VEGF expression in the knee articular cartilage of mice among the Sham, Dmm and Dmm+Th groups (n=4 in each group). (A) Immunohistochemistry staining of VEGF in the articular cartilage of the medial tibial plateau (magnification, x400, scale bar=100 µm). (B) Quantification of VEGF positive cells, based on the results of immunohistochemistry staining. The values are presented as the mean ± standard deviation. *P<0.05 compared with the Sham group; #P<0.05 compared with the Dmm group. Dmm, destabilization of the medial meniscus; Th, thalidomide; VEGF, vascular endothelial growth factor.

Article Snippet: An ELISA kit of VEGF (E-EL-M1292c) was purchased from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Immunohistochemical staining, Expressing, Immunohistochemistry, Staining, Standard Deviation

Journal: Molecular medicine reports

Article Title: Intraperitoneal injection of thalidomide alleviates early osteoarthritis development by suppressing vascular endothelial growth factor expression in mice.

doi: 10.3892/mmr.2018.8980

Figure Lengend Snippet: Figure 5. ELISA analysis of serum VEGF concentration of mice among the Sham, Dmm and Dmm+Th groups (n=8 in each group). The values are presented as the mean ± standard deviation. *P<0.05 compared with the Sham group; #P<0.05 compared with the Dmm group. Dmm, destabilization of the medial meniscus; Th, thalidomide; VEGF, vascular endothelial growth factor.

Article Snippet: An ELISA kit of VEGF (E-EL-M1292c) was purchased from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Standard Deviation

Journal: bioRxiv

Article Title: ASK1/p38-mediated NLRP3 inflammasome signaling pathway contributed to aberrant retinal angiogenesis in diabetic retinopathy

doi: 10.1101/763102

Figure Lengend Snippet: (A) The inflammatory related cytokine IL-6, TNF-α and IL-1β mRNA expression levels were enhanced in DR group. (B) The expression of vascular marker CD31 was increased in DR group by confocal detection. Meanwhile, (C) the VEGF secretion levels in DR group was increased compared with control group. Values are expressed as the mean ± SD (n = 6, *P < 0.05, **P < 0.01 and ***P < 0.001 compared with the control).

Article Snippet: The retinal tissue from control group, HG group and HG treated with inhibitor group were added in PBS and homogenized; the homogenate was centrifuged at 12000 g at 4°C for 10 min. Then, the supernatant was tested for VEGF by mouse VEGF ELISA kit (R&D Systems, MN, USA).

Techniques: Expressing, Marker, Control

Journal: bioRxiv

Article Title: ASK1/p38-mediated NLRP3 inflammasome signaling pathway contributed to aberrant retinal angiogenesis in diabetic retinopathy

doi: 10.1101/763102

Figure Lengend Snippet: (A) The protein expression level of NLRP3 was enhanced in DR groups; moreover, (B) blocking NLRP3, the mRNA expression of IL-6, TNF-α and IL-1β was down-regulated; meanwhile, (C) the vascular marker CD31 expression level and (D) VEGF secretion level were decreased through inhibiting NLRP3. In addition, in HRMEC cell model, (E) high glucose enhanced the protein expression of NLRP3, which was inhibited by NLPR3 inhibitor. (F) The inflammatory related cytokine IL-6, TNF-α and IL-1β was enhanced in HG-induced HRMEC cell model and was inhibited through blocking NLRP3. (G) The VEGF secretion level was enhanced, and was decreased after inhibiting NLRP3. Data are presented as the mean ± standard deviation from triplicate wells. *P < 0.05 and **P < 0.01 compared with the control. #P < 0.05 and ## P < 0.01 compared with the relative DR animal model group or HG-induced HRMEC cell group.

Article Snippet: The retinal tissue from control group, HG group and HG treated with inhibitor group were added in PBS and homogenized; the homogenate was centrifuged at 12000 g at 4°C for 10 min. Then, the supernatant was tested for VEGF by mouse VEGF ELISA kit (R&D Systems, MN, USA).

Techniques: Expressing, Blocking Assay, Marker, Standard Deviation, Control, Animal Model

Journal: bioRxiv

Article Title: ASK1/p38-mediated NLRP3 inflammasome signaling pathway contributed to aberrant retinal angiogenesis in diabetic retinopathy

doi: 10.1101/763102

Figure Lengend Snippet: (A) The protein expression levels of AKS1 and p38 were up-regulated in DR animal group. (B)The result of HG-induced HRMEC group was similar with DR animal group. Moreover, (C) the NLPR3, IL-6, TNF-α, IL-1β and VEGF protein expression levels were inhibited by (C) NLPR3 inhibitor and (D) p38 inhibitor.

Article Snippet: The retinal tissue from control group, HG group and HG treated with inhibitor group were added in PBS and homogenized; the homogenate was centrifuged at 12000 g at 4°C for 10 min. Then, the supernatant was tested for VEGF by mouse VEGF ELISA kit (R&D Systems, MN, USA).

Techniques: Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: An allosteric peptide inhibitor of HIF-1α regulates hypoxia-induced retinal neovascularization

doi: 10.1073/pnas.2017234117

Figure Lengend Snippet: Combination therapy of the CITED2 peptide and aflibercept rescues retinal neovascularization and vaso-obliteration in OIR. (A) Immunofluorescent staining of OIR retinas at P17 after intravitreal injection of aflibercept (200 ng), aflibercept and CITED2 peptide (200 ng aflibercept and 3.4 ng CITED2 peptide), or CITED2 peptide (3.4 ng) at P12. Retinal whole mounts were stained with GS-lectin. Representative images for each treatment are shown (Upper) and the same images are shown (Lower) with NV highlighted in red and VO highlighted in yellow as used for quantification. (Scale bars, 1 mm.) (B) Quantification of the percentage of VO area in whole retinas. (C) Quantification of the percentage of NV area in whole retinas. For B and C, n > 7 per group. P values were calculated using one-way ANOVA with Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (D) Validation of expression of HIF target genes in CITED2- and aflibercept-treated OIR retinas. Total RNA was isolated 24 h after intravitreal injection of CITED2 peptide (3.4 ng) or aflibercept (10 μg) on P12 (n = 6 per group). P values were calculated using multiple t tests. **P < 0.01, ***P < 0.001, ****P < 0.0001. (E) Quantification of VEGF protein levels in P15 retinas by ELISA (n = 6 per group). P values were calculated using one-way ANOVA with Tukey’s multiple comparisons test. *P < 0.05, ****P < 0.0001. For B–E, the mean and SEM are shown in red.

Article Snippet: Supernatants were then assayed without dilution in duplicate using the Mouse VEGF Quantikine ELISA kit (R&D Systems) according to the manufacturer’s protocol.

Techniques: Staining, Injection, Biomarker Discovery, Expressing, Isolation, Enzyme-linked Immunosorbent Assay

Journal: Antioxidants

Article Title: p38α MAPK-Mediated Redox Regulation of Transglutaminase 2 Drives Microvascular Leakage in Diabetic Retinas

doi: 10.3390/antiox15020192

Figure Lengend Snippet: p38 MAPK and TGase contribute to hyperglycemia-induced vascular leakage in HRECs and diabetic mouse retinas. ( A – F ) Diabetic C57BL/6 mice were intravitreally injected with 2 μL of 500 μmol/L SB203580 (SB) or 50 mmol/L cystamine (Cys) for 24 h. Transglutaminase (TGase) activity, vascular leakage, and vascular endothelial growth factor (VEGF) expression were assessed in the retinas. ( A ) Schematic illustration of diabetic mouse generation and in vivo experimental design. ( B , C ) Body weight ( B ) and blood glucose levels ( C ) were monitored weekly ( n = 6). ( D ) Vascular leakage visualized by fluorescein Isothiocyanate (FITC)-dextran angiography (scale bar, 100 μm) and fundus fluorescein angiography (scale bar, 300 μm). ( E ) VEGF expression (green) in retinal sections visualized by immunofluorescence with 4′,6-diamidino-2-phenylindole (DAPI) nuclear counterstain (blue). Scale bar, 100 μm. ( F ) Quantification of VEGF expression by ELISA ( n = 6). ( G ) In vitro endothelial monolayer permeability assay using 40-kDa FITC-dextran in human retinal endothelial cells (HRECs) ( n = 4). HRECs were incubated with 20 μmol/L SB or 50 μmol/L Cys and then treated with 10 ng/mL VEGF for 90 min. Statistical significance was determined using one-way ANOVA with Holm–Sidak’s multiple comparisons test. GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; NS, non-significant; *** p < 0.001.

Article Snippet: VEGF levels were determined using a commercial ELISA kit (MMV00; R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions [ ].

Techniques: Injection, Activity Assay, Expressing, In Vivo, Immunofluorescence, Enzyme-linked Immunosorbent Assay, In Vitro, Permeability, Incubation

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

Journal: Molecular Therapy. Nucleic Acids

Article Title: Nanoparticle delivery of VEGF-B mRNA promotes T cell infiltration within tumor and triggers robust antitumor immunity

doi: 10.1016/j.omtn.2025.102620

Figure Lengend Snippet: Characterization of VEGF-B mRNA modifications and mRNA-LNPs (A and B) IP-western blot analysis of VEGF-B protein expression in HEK293T and B16F10 cells after transfection of VEGF-B mRNA with different modifications and tailing method. Non, non-modified; m1Ψ, m1Ψ-modified and EPAP tailing; Ψ1, Ψ-modified and EPAP tailing; Ψ2, Ψ-modified and co-transcriptionally tailing. (C) Transmission electron microscopy observation of VEGF mRNA-LNP; scale bars, 200 μm. (D) Size distribution of VEGF mRNA-LNP and control mRNA-LNP detected by Malvern dynamic light scattering Zetasizer. (E) VEGF-B protein secretion levels in B16F10 cells transfected with VEGF-B mRNA-LNP, detected by ELISA. Cells in a six-well plate were transfected with 2 μg of the modified mRNA per well. Data are presented as the median ± standard error of the mean (SEM). Significance was determined with ordinary one-way ANOVA in (E). ∗∗∗∗ p < 0.0001.

Article Snippet: The concentration of VEGF-B protein was quantified through ELISA with the mouse VEGF-B ELISA kit (KOA0852, Rockland Immunochemicals Inc.), adhering strictly to the protocol provided by the manufacturer.

Techniques: Western Blot, Expressing, Transfection, Modification, Transmission Assay, Electron Microscopy, Control, Enzyme-linked Immunosorbent Assay