vegf protein concentration  (R&D Systems)

Journal: International Journal of Nanomedicine

Article Title: Combined Self-Assembled iRGD Polymersomes for Effective Targeted siRNA Anti-Tumor Therapy

doi: 10.2147/ijn.s383862

Figure Lengend Snippet: Figure 8 Antitumor effects of different siVEGF-delivery systems in nude mice bearing breast cancer via intravenous administration. (A) Images of mice on day 14 post administration. (B) Changes of animal weight with time. (C) Changes of tumor volume with time, *P < 0.01 vs saline only for data on day 14. (D) Relative VEGF protein level in tumor tissues (data from ELISA), *P < 0.01 vs saline. n = 5.

Article Snippet: The concentration VEGF protein in supernatants was determined using a Rat VEGF ELISA Kit (Boster) according to the manufacturer’s instructions.

Techniques: Saline, Enzyme-linked Immunosorbent Assay

Journal: Stem Cell Research & Therapy

Article Title: PTEN inhibitor VO-OHpic attenuates GC-associated endothelial progenitor cell dysfunction and osteonecrosis of the femoral head via activating Nrf2 signaling and inhibiting mitochondrial apoptosis pathway

doi: 10.1186/s13287-020-01658-y

Figure Lengend Snippet: VO-OHpic exerts protective effects on MPS-regulated angiogenesis of EPCs. a Representative images of tube formation of EPCs after treatment with 50-μM MPS or 50-μM MPS combined with 1-μM VO-OHpic for 48 h. b VEGF protein concentration in the culture medium of EPCs as determined by ELISA after treatment for 48 h. c Wound healing assay was performed and representative images were taken after 12 h and 24 h in EPCs treated with 50-μM MPS or 50-μM MPS combined with 1-μM VO-OHpic. d The wound closure rate was measured and relatively compared to the control group with no treatment of MPS and VO-OHpic. e VEGF, VEGFR1, and VEGFR2 protein levels were determined by western blot analysis after treatment for 48 h. f Band density ratios of VEGF, VEGFR1, and VEGFR2 to GAPDH in the western blot analysis were quantified by densitometry. All experiments were repeated for three times; ∗ P < 0.05 versus control, # P < 0.05 versus 50-μM MPS group. Error bar represents SD

Article Snippet: The vascular endothelial growth factor (VEGF) concentration was measured with a rat VEGF ELISA kit (Boster, China, EK0540).

Techniques: Protein Concentration, Enzyme-linked Immunosorbent Assay, Wound Healing Assay, Control, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: PTEN inhibitor VO-OHpic attenuates GC-associated endothelial progenitor cell dysfunction and osteonecrosis of the femoral head via activating Nrf2 signaling and inhibiting mitochondrial apoptosis pathway

doi: 10.1186/s13287-020-01658-y

Figure Lengend Snippet: Nrf2 activation is required for VO-OHpic-mediated anti-MPS effects in EPCs. a Relative mRNA expressions of Nrf2 as determined by RT-qPCR 2 days after siNrf2 transfection. b EPCs transfected with or without siNrf2 for 24 h were then treated with 50-μM MPS or 50-μM MPS combined with 1-μM VO-OHpic for 48 h. Nrf2, NQO-1, HO-1, and Trx protein levels were determined by western blot analysis. c Band density ratios of Nrf2, NQO-1, HO-1, and Trx to GAPDH in the western blots were quantified by densitometry. d The cleaved caspase 3 protein level was also determined by western blot analysis. e Band density ratios of cleaved caspase 3 to GAPDH in the western blots were quantified by densitometry. f Flow cytometric analysis of EPCs transfected with or without siNrf2 were stained with Annexin V-FITC/PI after treatment with 50-μM MPS or 50-μM MPS combined with 1-μM VO-OHpic for 48 h. g Percentage of apoptosis rates was expressed as means ± SD. h The VEGF protein level was determined by western blot analysis. i Band density ratios of VEGF to GAPDH in the western blots were quantified by densitometry. j Representative images of tube formation of EPCs in 12 h. EPCs were pre-transfected with and without siNrf2 for 24 h and then treated with 50-μM MPS combined with 1-μM VO-OHpic before seeded into Matrigel. k Representative images of EPCs transfected with or without siNrf2 with intracellular ROS stained by the fluorescence probe DCFH-DA after treatment for 48 h. l Flow cytometric analysis of ROS production after staining with DCFH-DA. m Bar graphs showing the mean fluorescence intensity (MFI) of ROS levels in EPCs. Data are shown as means ± SD. All experiments were repeated for three times; ∗ P < 0.05 versus EPCs without siNrf2 transfection. Error bar represents SD

Article Snippet: The vascular endothelial growth factor (VEGF) concentration was measured with a rat VEGF ELISA kit (Boster, China, EK0540).

Techniques: Activation Assay, Quantitative RT-PCR, Transfection, Western Blot, Staining, Fluorescence

Journal: Stem Cell Research & Therapy

Article Title: PTEN inhibitor VO-OHpic attenuates GC-associated endothelial progenitor cell dysfunction and osteonecrosis of the femoral head via activating Nrf2 signaling and inhibiting mitochondrial apoptosis pathway

doi: 10.1186/s13287-020-01658-y

Figure Lengend Snippet: Immunohistochemical staining of the sections of representative rat femoral heads. a Representative images of CD31 staining in the control, MPS, and MPS + VO-OHpic groups. b Relative CD31 expression was measured with the ImageJ software. ∗ P < 0.05 versus control, # P < 0.05 versus MPS group. Error bar represents SD. c Representative images of VEGF staining in the control, MPS, and MPS + VO-OHpic groups. d Relative VEGF expression was measured with the ImageJ software. ∗ P < 0.05 versus control, # P < 0.05 versus MPS group. Error bar represents SD. e Representative images of VEGFR2 staining in the control, MPS, and MPS + VO-OHpic groups. f Relative VEGFR2 expression was measured with the ImageJ software. g CD31-positive blood vessel number per field under × 200 magnification. h VEGFR2-positive blood vessel number per field under × 200 magnification. ∗ P < 0.05 versus control, # P < 0.05 versus MPS group. Error bar represents SD. Each group consisted of 10 rats/20 femoral heads ( N = 20)

Article Snippet: The vascular endothelial growth factor (VEGF) concentration was measured with a rat VEGF ELISA kit (Boster, China, EK0540).

Techniques: Immunohistochemical staining, Staining, Control, Expressing, Software

Journal: International Journal of Biological Sciences

Article Title: The role of monoamine oxidase A in HPV-16 E7-induced epithelial-mesenchymal transition and HIF-1α protein accumulation in non-small cell lung cancer cells

doi: 10.7150/ijbs.46966

Figure Lengend Snippet: MAOA knockout regulated HPV-16 E7-induced HIF-1α protein stability in NSCLC cells. (A) HIF-1α protein expression was analyzed by Western blotting. (B) VEGF protein concentration in the conditional media derived from stable A549 and NCI-H460 cells was determined by ELISA. (C) RT-qPCR analysis of VEGF mRNA expression in stable A549 and NCI-H460 cells. (D) The stable A549 and NCI-H460 cells (16 E7 and 16 E7-MAOA KO) were treated with MG132 (20 mmol/L) for 24 h, followed by analysis of HIF-1α protein expression. (E) The intracellular ROS level was determined by flow cytometry. All data are expressed as mean±SD of three independent experiments. * P < 0.05, ** P < 0.01.

Article Snippet: The stable-infected cells (Empty vector, 16 E7, and 16 E7-MAOA KO) were cultured for 24 h, and the VEGF protein concentration in the conditional media was measured using a human VEGF ELISA kit (Boster Biological Technology Co.Ltd.) according to the manufacturer's instructions.

Techniques: Knock-Out, Expressing, Western Blot, Protein Concentration, Derivative Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Flow Cytometry

Journal: International Journal of Biological Sciences

Article Title: The role of monoamine oxidase A in HPV-16 E7-induced epithelial-mesenchymal transition and HIF-1α protein accumulation in non-small cell lung cancer cells

doi: 10.7150/ijbs.46966

Figure Lengend Snippet: MAOA knockout inhibited HPV-16 E7-induced NSCLC growth, metastasis, and expression of EMT-related markers and HIF-1α proteins in vivo . The stable NCI-H460 cells (Empty vector, 16 E7, and 16 E7-MAOA KO) were respectively injected into subcutaneous (A-D) and intrapulmonary (E,F) of nude mice ( n = 8 mice/group). (A) The representative results of tumor growth. (B) The volume of subcutaneous xenograft tumors. (C) The weight of subcutaneous xenograft tumors. (D) Immunohistochemical staining results of Ki-67, MAOA, E-cadherin, N-cadherin, Slug, HIF-1α, and VEGF proteins in subcutaneous xenograft tumor tissues of nude mice. Scale bar =100 µm. (E) The sternal metastasis of NCI-H460 intrapulmonary tumors. (F) The rate of sternal metastasis. All data are expressed as mean±SD of three independent experiments. * P < 0.05, ** P < 0.01.

Article Snippet: The stable-infected cells (Empty vector, 16 E7, and 16 E7-MAOA KO) were cultured for 24 h, and the VEGF protein concentration in the conditional media was measured using a human VEGF ELISA kit (Boster Biological Technology Co.Ltd.) according to the manufacturer's instructions.

Techniques: Knock-Out, Expressing, In Vivo, Plasmid Preparation, Injection, Immunohistochemical staining, Staining