endothelial growth factor vegf concentration  (Boster Bio)

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