Journal: International Journal of Oncology

Article Title: Expression and epigenetic regulatory mechanism of BNIP3 in clear cell renal cell carcinoma

doi: 10.3892/ijo.2018.4603

Figure Lengend Snippet: BNIP3 expression in ccRCC tumor tissue samples, adjacent non-tumor tissue samples, and cell lines. (A) Relative protein expression levels of BNIP3, VHL, HIF-1α, and VEGF in tumor and adjacent non-tumor tissue samples from 30 cases of ccRCC were determined by WB using the levels of GAPDH as an internal control. Data are presented as the median and interquartile range. T, tumor tissues; N, adjacent non-tumor tissues. * P<0.05 compared with adjacent non-tumor tissues. Representative examples are shown. (B) RT-qPCR, demonstrating that BNIP3 mRNA expression was significantly lower in 786-O, ACHN, A498 and GRC-1-1 RCC cells (particularly 786-O cells) compared with normal renal HK-2 cells. BNIP3 mRNA expression levels were measured as percentages of that of HK-2. * P<0.05 compared with ACHN, GRC-1-1, 786-O, A498 cells. (C) BNIP3 protein levels in 786-O, ACHN, A498, GRC-1-1, and HK-2 cells were evaluated by WB, with GAPDH as a control. A representative example is shown. WB, western blotting; VHL, Hippel Lindau; HIF-1α, hypoxia-inducible factor-1α; VEGF, vascular endothelial growth factor; ccRCC, clear cell renal cell carcinoma; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: Corresponding membranes were then incubated with primary antibodies against BNIP3 (cat. no. B7931, 1:3,000; Sigma-Aldrich; now a brand of Merck, KGaA, Darmstadt, Germany), VHL (cat. no. 68547, 1:1,000; CST Biological Reagents Co., Ltd., Shanghai, China), HIF-1α (cat. no. 610959, 1:2,000; BD Biosciences, San Jose, CA, USA), VEGF (cat. no. BA0407, 1:100; Boster Biological Technology, Pleasanton, CA, USA), and GAPDH (cat. no. KC-5G4, 1:10,000; Kangchen BioTech Co., Ltd., Shanghai, China) at 4°C overnight.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Real-time Polymerase Chain Reaction

Journal: Bioscience Reports

Article Title: BMP9 exhibits dual and coupled roles in inducing osteogenic and angiogenic differentiation of mesenchymal stem cells

doi: 10.1042/BSR20201262

Figure Lengend Snippet: Primer oligonucleotide sequences used for PCR

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against vascular endothelial growth factor a (VEGFa; Abcam, Cambridge, U.S.A.), CD31 (Abcam, Cambridge, U.S.A.), and GAPDH (Bioworld Technology, MN, U.S.A.) at a dilution of 1:500 or 1:1000.

Techniques:

Journal: Bioscience Reports

Article Title: BMP9 exhibits dual and coupled roles in inducing osteogenic and angiogenic differentiation of mesenchymal stem cells

doi: 10.1042/BSR20201262

Figure Lengend Snippet: ( A ) BMP9-induced angiogenic differentiation markers expression in MSCs at mRNA level. Subconfluent iMEF cells were infected with AdBMP9 or AdGFP. At the indicated time points, total RNA of each group was isolated and subjected to RT-qPCR analysis with VEGFa ( a ), CD31 ( b ) primers. All samples were normalized with the reference gene GAPDH. Relative expression was calculated by dividing the relative expression values (i.e., gene/GAPDH) in AdBMP9 group with that from the AdGFP group. Each assay condition was done in triplicate. ‘*’ P <0.05, ‘**’ P <0.01, AdBMP9 group vs. AdGFP group. ( B ) BMP9-induced angiogenic differentiation marker expression in MSCs at protein levels. Subconfluent iMEF cells were infected with AdBMP9 or AdGFP. At the indicated time points, Western blot was used for detecting the expression of VEGFa ( a ) and CD31 ( c ). Relative protein expression levels were analyzed by Image Lab software using GAPDH as control ( b,d ). The results are expressed as mean ± SD of triplicate experiments, ‘**’ P <0.01, AdBMP9 group vs. AdGFP group.

Article Snippet: The PVDF membranes were incubated overnight at 4°C with primary antibodies against vascular endothelial growth factor a (VEGFa; Abcam, Cambridge, U.S.A.), CD31 (Abcam, Cambridge, U.S.A.), and GAPDH (Bioworld Technology, MN, U.S.A.) at a dilution of 1:500 or 1:1000.

Techniques: Expressing, Infection, Isolation, Quantitative RT-PCR, Marker, Western Blot, Software

Journal: Cancers

Article Title: Periostin Exon-21 Antibody Neutralization of Triple-Negative Breast Cancer Cell-Derived Periostin Regulates Tumor-Associated Macrophage Polarization and Angiogenesis

doi: 10.3390/cancers13205072

Figure Lengend Snippet: Human primer list.

Article Snippet: The membranes were incubated with primary antibodies against GAPDH (FUJIFILM, Japan, cat # 015–25473), periostin (Adipogen AG, San Diego, CA, USA, cat #AG-20B-0033-C100), Pn-21 Ab and secondary antibodies for ECL analysis.

Techniques:

Journal: Journal of Oncology

Article Title: Licoflavone A Suppresses Gastric Cancer Growth and Metastasis by Blocking the VEGFR-2 Signaling Pathway

doi: 10.1155/2022/5497991

Figure Lengend Snippet: LA arrested cells at G1 phase and induced apoptosis of VEGF-stimulated MKN-45 cells. (a) Flow cytometry assay of cell cycle distribution. (b) The quantitation of cell cycle distribution. The data are presented as mean ± SD ( n = 3). (c) The Hoechst 33342-stained nuclei were imaged under laser confocal microscopy. Bar = 50 μ m. Representative photographs of the morphological changes observed. The red arrow indicates apoptotic bodies and the white arrows indicate shrunken and deformed nuclei of apoptotic cells. (d) Apoptosis of LA-treated VEGF-stimulated MKN-45 cells was determined by flow cytometric analysis. (e) The quantitation of cell cycle distribution. The data are presented as mean ± SD ( n = 3). (f) A JC-10 kit was used to measure the mitochondrial membrane potential of VEGF-stimulated MKN-45 cells by laser confocal microscopy. CCCP staining results were used as the positive control. JC-10 is the monomer with green fluorescence, while JC-10 is the polymer with red fluorescence. Bar = 100 μ m. (g) Cyclin D1, c-Myc, Bcl-2, Bax, Cyt C, caspase 9, and cleaved-caspase 3 protein expression levels were detected by western blot.

Article Snippet: The primary antibodies to cyclin D1, c-Myc, E-cadherin, N-cadherin, Bcl-2, Bax, Cyt c, cleaved-caspase 3, PI3K, p-PI3K (Y607), AKT, p-AKT (Ser473), ERK1/2, p-ERK1/2 (Y204), GAPDH, and horseradish peroxidase- (HRP-) labeled goat anti-rabbit immunoglobulin G (IgG) (H+L) were purchased from ImmunoWay (Plano, TX).

Techniques: Flow Cytometry, Quantitation Assay, Staining, Confocal Microscopy, Membrane, Positive Control, Fluorescence, Polymer, Expressing, Western Blot