Journal: Scientific reports

Article Title: Characteristics of pulmonary microvascular structure in postnatal yaks.

doi: 10.1038/s41598-021-97760-z

Figure Lengend Snippet: Figure 4. Relative expression levels of VEGFA and EPAS1 in the plateau yak lung tissues at different developmental stages (1 day, 30 days, 180 days, adult). (A) Relative VEGFA mRNA levels in the lungs of plateau yaks at different developmental stages. (B) Relative EPAS1 mRNA levels in the lungs of plateau yaks at different developmental stages. (C) VEGFA protein expression at different developmental stages as determined by western blot analysis. (D) The graph represents a densitometric analysis of VEGFA protein expression. (Full- length blots are presented in Supplementary Figures S1 and S2). VEGFA protein content is expressed as relative levels and was quantified using ImagePro Plus software. VEGFA protein expression levels were normalized to β-Actin protein levels. All values above represent the mean ± SD (n = 3). Different letters (a, b, c, and d) indicate significant differences (P < 0.05).

Article Snippet: PVDF membranes were blocked in 10% nonfat (skimmed) milk for 3 h and then incubated in rabbit anti-VEGFA polyclonal antibody (OriGene, Maryland, USA) at 4 °C overnight.

Techniques: Expressing, Western Blot, Software

Journal: Oncology reports

Article Title: In vitro and in vivo activities of an antitumor peptide HM-3: A special dose-efficacy relationship on an HCT‑116 xenograft model in nude mice.

doi: 10.3892/or.2016.5077

Figure Lengend Snippet: Figure 8. Percent of VEGF expressing cells on the sections of tumor tis- sues from different groups. (A) Typical image for section from the control group. (B) Typical image for section from sunitinib (60 mg/kg) treatment group. (C) Typical image for section from HM-3 (3 mg/kg) treatment group. (D) Typical image for section from HM-3 (48 mg/kg) treatment group. (E) Comparison of the percent of VEGF-staining positive cells in dif- ferent groups. Mean ± SD, *P<0.05, compared with the control group.

Article Snippet: Tumors were immediately fixed in 4% formaldehyde and embedded in paraffin, then, sectioned for immunohistochemical staining of CD31, HIF-1α and VEGF with rabbit anti-CD31, HIF-1α and VEGF polyclonal antibodies (Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.).

Techniques: Expressing, Control, Comparison, Staining

Journal: Frontiers in Immunology

Article Title: Adipocytes Promote Early Steps of Breast Cancer Cell Dissemination via Interleukin-8

doi: 10.3389/fimmu.2018.01767

Figure Lengend Snippet: Addition of breast adipocytes (BAd) significantly increased interleukin-8 (IL-8) secretion but decreased vascular endothelial growth factor (VEGF) secretion compared to breast cancer cells (BCC) cultured alone and anti-IL-8 and anti-VEGF significantly decreased BAd-induced angiogenesis in primary tumors in zebrafish. BCC were cultured in 3D spheres alone or in combination with BAd. Secreted IL-8 and VEGF were analyzed as described in Section “ .” Prior injections, breast pre-adipocytes were differentiated for 12 days and estrogen receptor positive (ER+) BCC were cultured ± β-estradiol (E2) 1 nM for 48 h. All BCC were labeled with 4 µg/ml Fast DiI™ oil red dye. Cells were injected ± anti-IL-8, anti-VEGF, or isotype control at 0.1 mg/ml ± E2 1 nM into the perivitelline space of 2 days old zebrafish embryos, which expressed enhanced green fluorescent protein in endothelial cells. (A) BAd mammospheres alone and low metastatic ER+ MCF-7 ± 90% BAd mammospheres were cultured ± E2 1 nM during 7 days, n = 4–5 in each group. (B) BAd mammospheres alone an ER+ T47D with intrinsically higher metastatic capacity ± 90% BAd mammospheres were cultured ± E2 1 nM during 7 days, n = 5–6 in each group. (C) Estrogen receptor negative (ER−) metastatic MDA-MB-23 ± 90% BAd and BAd mammospheres were cultured during 7 days, n = 4–5 in each group. (D) MCF-7 cells were injected alone or in combination with 50% BAd ± E2 1 nM, tumor angiogenesis was analyzed 3 days post-injections, n = 12–18 in each group. (E) T47D cells were injected alone or in combination with 50% Bad ± E2 1 nM, tumor angiogenesis was analyzed 3 days post-injections, n = 10 in each group. (F) MDA-MB-231 cells were injected alone or in combination with 50% BAd, tumor angiogenesis was analyzed 3 days post-injections, n = 7–10 in each group. Representative confocal images are shown for each cell line. BV = blood vessels. Results are presented as mean ± SEM and analyzed by Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Data are representative of at least two independent experiments.

Article Snippet: After 24 h co-culture, treatment ± E2 1 nM, rabbit anti-human IL-8, rabbit anti-human VEGF (Acris Antibodies GmbH Cat # PP1073P1, RRID:AB_1008432), or rabbit isotype antibodies at 1 μg/ml was performed for 3 days.

Techniques: Cell Culture, Labeling, Injection, Control

Journal: Frontiers in Immunology

Article Title: Adipocytes Promote Early Steps of Breast Cancer Cell Dissemination via Interleukin-8

doi: 10.3389/fimmu.2018.01767

Figure Lengend Snippet: Anti-interleukin-8 (IL-8) treatment significantly decreased breast cancer cells (BCC) dissemination induced by breast adipocytes (BAd). Prior injections, breast pre-adipocytes were differentiated for 12 days and estrogen receptor positive (ER+) BCC were cultured ± β-estradiol (E2) 1 nM for 48 h. All BCC were labeled with 4 µg/ml Fast DiI™ oil red dye. Cells were injected ± anti-IL-8, anti-VEGF, or isotype control at 0.1 mg/ml ± E2 1 nM into the perivitelline space of 2 days old zebrafish embryos, which expressed enhanced green fluorescent protein in endothelial cells. (A) MCF-7 cells were injected alone or in combination with 50% BAd ± E2 1 nM. BCC dissemination was evaluated 3 days post-injections, n = 18–27 in each group. (B) T47D cells were injected alone or in combination with 50% BAd ± E2 1 nM. BCC dissemination was evaluated 3 days post-injections, n = 16–28 in each group. (C) MDA-MB-231 cells were injected alone or in combination with 50% BAd. BCC dissemination was evaluated 3 days post-injections, n = 14–18 in each group. (D) MCF-7, T47D, and MDA-MB-231 cells were cultured alone or in combination with 50% breast pre-adipocytes in vitro during 24 h, and migration of cells was determined as described in Section “ ,” n = 6 in each group. BV = blood vessels. Arrows indicate disseminated BCC. Results are presented as mean ± SEM and analyzed by Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001. Data are representative of at least two independent experiments.

Article Snippet: After 24 h co-culture, treatment ± E2 1 nM, rabbit anti-human IL-8, rabbit anti-human VEGF (Acris Antibodies GmbH Cat # PP1073P1, RRID:AB_1008432), or rabbit isotype antibodies at 1 μg/ml was performed for 3 days.

Techniques: Cell Culture, Labeling, Injection, Control, In Vitro, Migration

Journal: Frontiers in Immunology

Article Title: Adipocytes Promote Early Steps of Breast Cancer Cell Dissemination via Interleukin-8

doi: 10.3389/fimmu.2018.01767

Figure Lengend Snippet: Anti-interleukin-8 (αIL-8) decreased vascular endothelial growth factor (VEGF) and CCL5 secretion, which affected estrogen receptor positive (ER+) breast cancer cells (BCC) dissemination. For monolayer co-cultures, breast pre-adipocytes were differentiated for 5 days before ER+ BCC were added at 4 × 10 3 cells/well. For zebrafish experiments, breast pre-adipocytes were differentiated for 12 days, MCF-7 cells were cultured + β-estradiol (E2) 1 nM for 48 h and labeled with 4 µg/ml Fast DiI™ oil red dye before injected into the perivitelline space of 2 days old zebrafish embryos, which expressed enhanced green fluorescent protein in endothelial cells. (A) MCF-7 cells were co-cultured with 50% breast adipocytes (BAd) in the presence or absence of αIL-8, anti-VEGF (αVEGF), or control isotype (Iso) antibodies at 1 µg/ml during 3 days in the presence of E2 1 nM, and secreted cytokines were quantified as described in Section “ ,” n = 5–4 in each group. (B) T47D cells were co-cultured with 50% BAd in the presence or absence of αIL-8, αVEGF, or control Iso antibodies at 1 µg/ml during 3 days in the presence of E2 1 nM, and secreted cytokines were quantified as described in Section “ ,” n = 6–5 in each group. (C) MCF-7 cells were injected in zebrafish embryos alone or in combination with 50% BAd ± anti-CCL5 (αCCL5) or Iso control antibody at 0.1 mg/ml and E2 1 nM, as described in Section “ .” MCF-7 dissemination was analyzed 3 days post-injections, n = 13–27 in each group. Representative images of zebrafish embryos are shown. Arrows show disseminated BCC cells. BV = blood vessels. Results are presented as mean ± SEM, Student’s t -test, * p < 0.05, *** p < 0.001. Data are representative of at least two independent experiments.

Article Snippet: After 24 h co-culture, treatment ± E2 1 nM, rabbit anti-human IL-8, rabbit anti-human VEGF (Acris Antibodies GmbH Cat # PP1073P1, RRID:AB_1008432), or rabbit isotype antibodies at 1 μg/ml was performed for 3 days.

Techniques: Cell Culture, Labeling, Injection, Control

Journal: Journal of traditional Chinese medicine = Chung i tsa chih ying wen pan

Article Title: Combinatorial effects of naomai yihao capsules and vascular endothelial growth factor gene-transfected bone marrow mesenchymal stem cells on angiogenesis in cerebral ischemic tissues in rats.

doi: 10.1016/s0254-6272(12)60038-7

Figure Lengend Snippet: Figure 1 CD34 expression in the brain of rats among groups on day 21 (SABC staining, x400). A: Model group, B: BMSC group, C: BMSC/VEGF group, D: NMYH group, E: BMSC/VEGF+NMYH group.

Article Snippet: The providers of reagents, materials and instruments are as follows: pEGFP-VEGF165 plasmid, the Stem Cell Research Center of Sun Yat-sen University (Guangzhou, China); Lipofectamine 2000, Invitrogen Co Ltd.(Grand Island, NY, USA); plasmid extraction kits, Tiangen Biotech (Beijing) Co Ltd. (Beijing, China); rabbit anti-rat VEGF antibody (Lot No. BS-0279R) and rabbit anti-rat CD34 antibody (Lot No. BS-0646R), Beijing Biosynthesis Biotechnology Co Ltd. (Beijing, China); SABC (Lot No. SP-9001) and DAB (Lot No. ZLI-9032) kits, Zhongshan Goldenbridge Biotechnology Co Ltd. (Beijing, China); fluorescein isothiocyanate-labeled dextran (FITC-Dextran; molecular weight: 2 × 106), Sigma-Aldrich Co., (St. Louis, MO, USA); laser scanning confocal microscope (LSCM), LEICA Microsystems Ltd., (Wetzlar, Germany); and Image-Pro Plus 6.0 image analysis software (Model No. 41M60032-00032), Media Cybernetics Inc., (Bethesda, MD, USA).

Techniques: Expressing, Staining

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: linc00958/miR-185-5p/RSF-1 modulates cisplatin resistance and angiogenesis through AKT1/GSK3β/VEGFA pathway in cervical cancer

doi: 10.1186/s12958-022-00995-2

Figure Lengend Snippet: Antibodies used in western blot and IHC experiments

Article Snippet: The primary antibody against VEGFA (bs-4572R, Bioss, Beijing, China) was diluted with 1% donkey serum (1:200) and then added in each well.

Techniques: Western Blot