rabbit polyclonal antibody against vegfa  (Wanleibio)

Journal: Renal Failure

Article Title: miR-29a-3p/ Vegfa axis modulates high phosphate-induced vascular smooth muscle cell calcification

doi: 10.1080/0886022x.2025.2489712

Figure Lengend Snippet: Figure 1. Identification of Vegfa as a direct target of miR-29a-3p in VSMCs using bioinformatics and dual-luciferase assay. (A) Venn diagram depicting the overlap of miR-29a-3p target genes predicted by four miRNA target prediction tools: miRTarBase, miRWalk, TargetScan, and miRDB. The box highlights the seven genes identified as common targets across all tools. (B) Sequence alignment of miR-29a-3p with the 3′-UTR region of wild-type and mutated Vegfa mRNA. The predicted binding site and seed region are marked in red. (C) Dual-luciferase reporter assay results for VSMCs co-transfected with a control vector, a vector containing the wild type (WT) Vegfa 3′-UTR sequence, or the MUT Vegfa sequence, along with miR-29a-3p mimics or a negative control miRNA (NT). Relative luciferase activity was measured, and data are presented as mean ± SD from three independent experiments.

Article Snippet: VSMCs recovered by centrifugation were homogenized and separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (China National Pharmaceutical Group Corporation Chemical reagent Co., Ltd., China; Cat. #30166428). the proteins were then transferred to polyvinylidene fluoride membranes (Millipore Co., uSa; Cat. #ISEQ00010). the blots were stained with rabbit polyclonal antibodies against VEGFa (1:10,000; Proteintech, uSa; Cat. #19003-1-aP), and mouse monoclonal antibodies against β-actin (1:10,000, Servicebio, China; Cat. #Gb15001), followed by anti-rabbit IgG (1:10,000, SIMubIotECH, China; Cat. #S2001), and anti-mouse IgG horseradish peroxidase conjugates (1:10,000, abclonal, China; Cat. #aS003).

Techniques: Luciferase, Sequencing, Binding Assay, Reporter Assay, Transfection, Control, Plasmid Preparation, Negative Control, Activity Assay

Journal: Renal Failure

Article Title: miR-29a-3p/ Vegfa axis modulates high phosphate-induced vascular smooth muscle cell calcification

doi: 10.1080/0886022x.2025.2489712

Figure Lengend Snippet: Figure 2. miR-29a-3p overexpression suppresses Vegfa expression in VSMCs. (A) Representative Western blot analysis of VEGFA protein levels in VSMCs under standard culture conditions (control), high phosphate conditions (Pi), and high phosphate conditions following miR-29a-3p overexpression (Pi + miR-29a-3p). β-actin served as a loading control. Quantitative analysis of VEGFA protein levels is shown as relative fold change normalized to β-actin. (B) Vegfa mRNA levels quantified using RT-qPCR in the same experimental groups. GAPDH was used as an internal control for normalization. Data represent the mean ± SD of five independent biological replicates.

Article Snippet: VSMCs recovered by centrifugation were homogenized and separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (China National Pharmaceutical Group Corporation Chemical reagent Co., Ltd., China; Cat. #30166428). the proteins were then transferred to polyvinylidene fluoride membranes (Millipore Co., uSa; Cat. #ISEQ00010). the blots were stained with rabbit polyclonal antibodies against VEGFa (1:10,000; Proteintech, uSa; Cat. #19003-1-aP), and mouse monoclonal antibodies against β-actin (1:10,000, Servicebio, China; Cat. #Gb15001), followed by anti-rabbit IgG (1:10,000, SIMubIotECH, China; Cat. #S2001), and anti-mouse IgG horseradish peroxidase conjugates (1:10,000, abclonal, China; Cat. #aS003).

Techniques: Over Expression, Expressing, Western Blot, Control, Quantitative RT-PCR

Journal: Renal Failure

Article Title: miR-29a-3p/ Vegfa axis modulates high phosphate-induced vascular smooth muscle cell calcification

doi: 10.1080/0886022x.2025.2489712

Figure Lengend Snippet: Figure 3. Vegfa knockdown and miR-29a-3p overexpression mitigate high phosphate-induced VSMCs calcification. (A–C) Analysis of Vegfa knockdown effects in VSMCs cultured in normal media (control) or high phosphate conditions (Pi). (A) Vegfa mRNA levels were quantified via RT-qPCR. (B) ARS with corresponding spectrophotometric quantification to assess cellular mineralization. (C) Quantification of intracellular Ca2+ content. Cells were treated with small interfering RNA targeting Vegfa (siVegfa) or a NT control. (D–F) Investigation of the role of miR-29a-3p in VSMCs cultured under high phosphate conditions. (D) Vegfa mRNA expression measured using RT-qPCR, (E) mineralization assessed using ARS and spectrophotometric quantification, and (F) intracellular Ca2+ levels were quantified. Cells were transfected with either an empty vector (VE) or a Vegfa-overexpressing construct (OE), combined with a NT miRNA mimic or a miR-29a-3p mimic. All experiments were performed in five replicates, and error bars represent the SD. GAPDH was used as an internal control in RT-qPCR experiments. Scale bar = 100 µm.

Article Snippet: VSMCs recovered by centrifugation were homogenized and separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (China National Pharmaceutical Group Corporation Chemical reagent Co., Ltd., China; Cat. #30166428). the proteins were then transferred to polyvinylidene fluoride membranes (Millipore Co., uSa; Cat. #ISEQ00010). the blots were stained with rabbit polyclonal antibodies against VEGFa (1:10,000; Proteintech, uSa; Cat. #19003-1-aP), and mouse monoclonal antibodies against β-actin (1:10,000, Servicebio, China; Cat. #Gb15001), followed by anti-rabbit IgG (1:10,000, SIMubIotECH, China; Cat. #S2001), and anti-mouse IgG horseradish peroxidase conjugates (1:10,000, abclonal, China; Cat. #aS003).

Techniques: Knockdown, Over Expression, Cell Culture, Control, Quantitative RT-PCR, Small Interfering RNA, Expressing, Transfection, Plasmid Preparation, Construct

Journal: Molecular Cancer

Article Title: MicroRNA-29c functions as a tumor suppressor by targeting VEGFA in lung adenocarcinoma

doi: 10.1186/s12943-017-0620-0

Figure Lengend Snippet: MiR-29c modulates LAD cells through targeting VEGFA. a The VEGFA 3’-UTR containing the wildtype or mutant miR-29c binding sequence was inserted into downstream of the luciferase reporter vector. The mutated sequences are in italic. b The dual luciferase reporter assay revealed that the luciferase activity controlled by VEGFA 3’-UTR was inhibited by ectopic miR-29c expression in A549 cells. *** p < 0.001. c VEGFA mRNA levels determined by qRT-PCR following treatment of A549 cells with miR-29c mimic or Anip973 cells with miR-29c inhibitor. ** p < 0.01. d ELISA assay of secreted VEGFA protein levels in TCM after treatment of A549 cells with miR-29c mimic or Anip973 cells with miR-29c inhibitor. * p < 0.05, ** p < 0.01. e Western blot analyses of VEGFA protein levels following treatment of A549 cells with miR-29c mimic or Anip973 cells with miR-29c inhibitor. β-actin was used as control

Article Snippet: After blocking with 3% bovine serum albumin (BSA) for 1 h, the membranes were then incubated with primary antibodies overnight at 4 °C: a rabbit polyclonal antibody against VEGFA (1:500, Proteintech, China) and a mouse monoclonal antibody for β-actin (1:5000, Santa Cruz, USA).

Techniques: Mutagenesis, Binding Assay, Sequencing, Luciferase, Plasmid Preparation, Reporter Assay, Activity Assay, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Molecular Cancer

Article Title: MicroRNA-29c functions as a tumor suppressor by targeting VEGFA in lung adenocarcinoma

doi: 10.1186/s12943-017-0620-0

Figure Lengend Snippet: Forced expression of VEGFA abolishes the phenotype created by mimic transfection in A549 cells. a and b Cell migration and invasion abilities were determined in A549 cells transfected with miR-29c mimic, mimic-NC and miR-29c mimic plus VEGFA expression plasmid. *** p < 0.001. c Cell proliferation abilities were determined in A549 cells transfected with miR-29c mimic, mimic-NC and miR-29c mimic plus VEGFA expression plasmid. * p < 0.05, ** p < 0.01. d and e HUVECs were cultured in TCM derived from A549 cells transfected with miR-29c mimic, mimic-NC and miR-29c mimic plus VEGFA expression plasmid. Representative images of tube formation and the relative number of tube branches measured in random 10 photographic fields are presented. ** p < 0.01, *** p < 0.001

Article Snippet: After blocking with 3% bovine serum albumin (BSA) for 1 h, the membranes were then incubated with primary antibodies overnight at 4 °C: a rabbit polyclonal antibody against VEGFA (1:500, Proteintech, China) and a mouse monoclonal antibody for β-actin (1:5000, Santa Cruz, USA).

Techniques: Expressing, Transfection, Migration, Plasmid Preparation, Cell Culture, Derivative Assay

Journal: Molecular Cancer

Article Title: MicroRNA-29c functions as a tumor suppressor by targeting VEGFA in lung adenocarcinoma

doi: 10.1186/s12943-017-0620-0

Figure Lengend Snippet: Correlation of miR-29c with MVD and VEGFA expression. a Correlation analysis between miR-29c and MVD. Red line represents linear regression line. b miR-29c expression levels in different VEGFA expression groups

Article Snippet: After blocking with 3% bovine serum albumin (BSA) for 1 h, the membranes were then incubated with primary antibodies overnight at 4 °C: a rabbit polyclonal antibody against VEGFA (1:500, Proteintech, China) and a mouse monoclonal antibody for β-actin (1:5000, Santa Cruz, USA).

Techniques: Expressing

Journal: PLoS ONE

Article Title: ZEB1 Upregulates VEGF Expression and Stimulates Angiogenesis in Breast Cancer

doi: 10.1371/journal.pone.0148774

Figure Lengend Snippet: Genes regulated by ZEB1.

Article Snippet: The following antibodies were used: goat polyclonal Ab against ZEB1 (ab81972; Abcam) at dilution of 1:1000, rabbit polyclonal Ab against VEGFA (19003-1-AP; Proteintech) at dilution of 1:1000, rabbit polyclonal Ab against VEGFC (22601-1-AP; Proteintech) at dilution of 1:500, and mouse monoclonal Ab against actin (A-4700; Sigma) at dilution of 1:1000.

Techniques: Control

Journal: PLoS ONE

Article Title: ZEB1 Upregulates VEGF Expression and Stimulates Angiogenesis in Breast Cancer

doi: 10.1371/journal.pone.0148774

Figure Lengend Snippet: (A) MDA-MB-231 cells were transiently transfected with human ZEB1 expression plasmid or empty vector control. At the indicated time points, expression of ZEB1, VEGFA and VEGFC was verified by Western blotting. Actin was used as a loading control. (B) Production of VEGFA and VEGFC protein were verified by ELISA at the indicated time points following ZEB1 overexpression. * P < 0.05, ** P < 0.01 vs respective control in one-way ANOVA followed by Tukey’s Honestly Significant Difference test. (C) HUVECs cultured in the presence or absence of VEGFA (20 and 40 ng/mL) or anti-VEGFA neutralized Ab (1 μg/mL) along with ZEB1/231-derived conditioned medium were subjected to a tube formation assay and photographed. (D) Quantification of tube formation was expressed as length of capillary tubes formed per mm 2 . * P < 0.05, ** P < 0.01 vs respective control in Student’s t -test.

Article Snippet: The following antibodies were used: goat polyclonal Ab against ZEB1 (ab81972; Abcam) at dilution of 1:1000, rabbit polyclonal Ab against VEGFA (19003-1-AP; Proteintech) at dilution of 1:1000, rabbit polyclonal Ab against VEGFC (22601-1-AP; Proteintech) at dilution of 1:500, and mouse monoclonal Ab against actin (A-4700; Sigma) at dilution of 1:1000.

Techniques: Transfection, Expressing, Plasmid Preparation, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Over Expression, Cell Culture, Derivative Assay, Tube Formation Assay

Journal: PLoS ONE

Article Title: ZEB1 Upregulates VEGF Expression and Stimulates Angiogenesis in Breast Cancer

doi: 10.1371/journal.pone.0148774

Figure Lengend Snippet: Positive correlation between ZEB1 and VEGFA expression in breast cancer.

Article Snippet: The following antibodies were used: goat polyclonal Ab against ZEB1 (ab81972; Abcam) at dilution of 1:1000, rabbit polyclonal Ab against VEGFA (19003-1-AP; Proteintech) at dilution of 1:1000, rabbit polyclonal Ab against VEGFC (22601-1-AP; Proteintech) at dilution of 1:500, and mouse monoclonal Ab against actin (A-4700; Sigma) at dilution of 1:1000.

Techniques: Expressing

Journal: PLoS ONE

Article Title: ZEB1 Upregulates VEGF Expression and Stimulates Angiogenesis in Breast Cancer

doi: 10.1371/journal.pone.0148774

Figure Lengend Snippet: The non-negative percentage analysis for VEGFA (A) and CD31 (B) indicates a positive correlation with ZEB1 expression in breast cancer tumors from 228 subjects. (C) Representative images of immunohistochemical staining of ZEB1, VEGFA, and CD31 in tumors from 4 cases are shown. Scale bars, 25 μm.

Article Snippet: The following antibodies were used: goat polyclonal Ab against ZEB1 (ab81972; Abcam) at dilution of 1:1000, rabbit polyclonal Ab against VEGFA (19003-1-AP; Proteintech) at dilution of 1:1000, rabbit polyclonal Ab against VEGFC (22601-1-AP; Proteintech) at dilution of 1:500, and mouse monoclonal Ab against actin (A-4700; Sigma) at dilution of 1:1000.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: ZEB1 Upregulates VEGF Expression and Stimulates Angiogenesis in Breast Cancer

doi: 10.1371/journal.pone.0148774

Figure Lengend Snippet: MDA-MB-231 cells were transiently transfected with the human ZEB1 expression plasmid or empty vector control, followed by treatment with PI-103 (10 μM) or SB203580 (10 μM). At the indicated time points, upregulation of VEGFA expression were verified by qPCR (A and B), Western blotting (C and D) and ELISA (E and F) in ZEB1-expressing versus control cells. GAPDH and actin were used to normalize VEGFA levels. * P < 0.05, ** P < 0.01 vs respective control in Student’s t -test.

Article Snippet: The following antibodies were used: goat polyclonal Ab against ZEB1 (ab81972; Abcam) at dilution of 1:1000, rabbit polyclonal Ab against VEGFA (19003-1-AP; Proteintech) at dilution of 1:1000, rabbit polyclonal Ab against VEGFC (22601-1-AP; Proteintech) at dilution of 1:500, and mouse monoclonal Ab against actin (A-4700; Sigma) at dilution of 1:1000.

Techniques: Transfection, Expressing, Plasmid Preparation, Control, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: ZEB1 Upregulates VEGF Expression and Stimulates Angiogenesis in Breast Cancer

doi: 10.1371/journal.pone.0148774

Figure Lengend Snippet: (A) Sequential deletion and mutation of SP1 elements on the human VEGFA promoter were fused to the luciferase reporter. (B) MDA-MB-231 cells were co-transfected with the ZEB1 expression plasmid (1 μg/well) and different wild-type VEGFA promoter luciferase reporter constructs (1 μg/well). Extract luciferase activities were determined 48 h after transfection using a Betascope analyzer. Luciferase values were normalized to Renilla activities. * P < 0.05 vs respective control in Student’s t -test. (C) MDA-MB-231 cells were co-transfected with ZEB1 expression plasmid and wild-type or mutant VEGFA promoter luciferase reporters. Extract luciferase activities were determined 48 h after transfection using a Betascope analyzer. Luciferase values are normalized with Renilla activities. * P < 0.05 vs respective control in Student’s t -test. (D) The association of SP1 with the proximal human VEGFA promoter was analyzed by ChIP assay, using polyclonal Ab against SP1 or unrelated IgG Ab. The amplified sequence of the VEGFA promoter fragment containing SP1 elements is shown. Input DNA amounts were confirmed by equal loading of chromatin. (E) Overexpression of ZEB1 significantly enhanced the recruitment of SP1 to the endogenous VEGFA promoter as confirmed by a quantitative ChIP assay. * P < 0.05 vs respective control in Student’s t -test.

Article Snippet: The following antibodies were used: goat polyclonal Ab against ZEB1 (ab81972; Abcam) at dilution of 1:1000, rabbit polyclonal Ab against VEGFA (19003-1-AP; Proteintech) at dilution of 1:1000, rabbit polyclonal Ab against VEGFC (22601-1-AP; Proteintech) at dilution of 1:500, and mouse monoclonal Ab against actin (A-4700; Sigma) at dilution of 1:1000.

Techniques: Mutagenesis, Luciferase, Transfection, Expressing, Plasmid Preparation, Construct, Control, Amplification, Sequencing, Over Expression