Journal: Current Issues in Molecular Biology

Article Title: IGFBP5 Promotes Atherosclerosis in APOE −/− Mice Through Phenotypic Transformation of VSMCs

doi: 10.3390/cimb47070555

Figure Lengend Snippet: Overexpression of IGFBP5 does not influence food consumption by and the body weight of ApoE −/− mice. ( A ) IGFBP5 mRNA expression level in skeletal muscle of ApoE −/− mice at 10 weeks of experimental treatment ( n = 3). ( B ) Plasma IGFBP5 protein levels at 10 weeks of experimental treatment ( n = 3). ( C ) Weight change curve ( n = 15). ( D ) Comparison of average daily food intake during the entire experimental treatment among groups ( n = 15). Data are expressed as the mean ± SEM. * p < 0.05. NS means no significant difference.

Article Snippet: The primary antibody used in this experiment was a rabbit anti- IGFBP5 antibody (1:500, #55205-1-AP; Proteintech Group, Chicago, IL, USA).

Techniques: Over Expression, Expressing, Clinical Proteomics, Comparison

Journal: Current Issues in Molecular Biology

Article Title: IGFBP5 Promotes Atherosclerosis in APOE −/− Mice Through Phenotypic Transformation of VSMCs

doi: 10.3390/cimb47070555

Figure Lengend Snippet: Overexpression of IGFBP5 enhances the development of atherosclerotic plaques in ApoE −/− mice. Oil Red O staining of the aorta ( A ) and statistical quantification ( B ). ( C ) H&E staining (up panel) and Oil Red O staining (down panel) of cardiac outflow tract sections. Scale bar: 500 μm. ( D ) Statistical quantification of neutral lipids in the plaques of the cardiac outflow tract sections stained with Oil Red O ( n = 10). Data are expressed as the mean ± SEM. * p < 0.05. NS means no significant difference.

Article Snippet: The primary antibody used in this experiment was a rabbit anti- IGFBP5 antibody (1:500, #55205-1-AP; Proteintech Group, Chicago, IL, USA).

Techniques: Over Expression, Staining

Journal: Current Issues in Molecular Biology

Article Title: IGFBP5 Promotes Atherosclerosis in APOE −/− Mice Through Phenotypic Transformation of VSMCs

doi: 10.3390/cimb47070555

Figure Lengend Snippet: Effect of overexpression of IGFBP5 on blood lipid levels in ApoE −/− mice. Plasma was collected from each group of mice ( n = 15) at week 0, 4, 8, and 12 of treatment to measure total cholesterol (TC) ( A ), total triglycerides (TG) ( B ), low-density lipoprotein cholesterol (LDL-C) ( C ), and high-density lipoprotein cholesterol (HDL-C) ( D ) levels. Data are expressed as the mean ± SEM.

Article Snippet: The primary antibody used in this experiment was a rabbit anti- IGFBP5 antibody (1:500, #55205-1-AP; Proteintech Group, Chicago, IL, USA).

Techniques: Over Expression, Clinical Proteomics

Journal: Current Issues in Molecular Biology

Article Title: IGFBP5 Promotes Atherosclerosis in APOE −/− Mice Through Phenotypic Transformation of VSMCs

doi: 10.3390/cimb47070555

Figure Lengend Snippet: Transcriptome analysis of aortic tissues in ApoE −/− mice following overexpression of IGFBP5 . ( A , B ) Heatmap representing global gene expression in aortic tissues of AAV-GFP and AAV- IGFBP5 mice after 12 weeks of being fed a HFD ( n = 4).

Article Snippet: The primary antibody used in this experiment was a rabbit anti- IGFBP5 antibody (1:500, #55205-1-AP; Proteintech Group, Chicago, IL, USA).

Techniques: Over Expression, Gene Expression

Journal: Current Issues in Molecular Biology

Article Title: IGFBP5 Promotes Atherosclerosis in APOE −/− Mice Through Phenotypic Transformation of VSMCs

doi: 10.3390/cimb47070555

Figure Lengend Snippet: Gene Ontology analyses of the differentially expression genes in aortic tissues in ApoE −/− mice following overexpression of IGFBP5 . ( A ) Gene Ontology analyses of the differentially expression genes in aortic of AAV-GFP and AAV- IGFBP5 mice after 12 weeks of HFD ( n = 4). ( B ) GO analysis was performed based on three categories: biological process (BP), molecular function (MF), and cellular component (CC).

Article Snippet: The primary antibody used in this experiment was a rabbit anti- IGFBP5 antibody (1:500, #55205-1-AP; Proteintech Group, Chicago, IL, USA).

Techniques: Expressing, Over Expression

Journal: Current Issues in Molecular Biology

Article Title: IGFBP5 Promotes Atherosclerosis in APOE −/− Mice Through Phenotypic Transformation of VSMCs

doi: 10.3390/cimb47070555

Figure Lengend Snippet: Overexpression of IGFBP5 induces VSMCs to adopt a proliferative state. ( A ) Flow cytometric analysis of cell cycle distribution of immortalized mouse vascular smooth muscle cell line treated with different concentrations of IGFBP5 during proliferation ( n = 3). ( B ) Cell cycle distribution statistics ( n = 3). ( C ) The CCK-8 kit was used to detect the effect of different concentrations of IGFBP5 on VSMC proliferation for 24 h ( n = 5). ( D ) Effects of different concentrations of IGFBP5 on cell migration in cell scratch assays ( n = 3). ( E ) Statistics of relative distance of cell migration ( n = 3). ( F ) Real-time qPCR analysis of ACTA2 , MYH11 , TAGLN , CNN1 and KLF4 transcription expression in immortalized mouse vascular smooth muscle cells treated with different concentrations of IGFBP5 ( n = 3). GAPDH was used as a control. Data are expressed as the mean ± SEM. * p < 0.05.

Article Snippet: The primary antibody used in this experiment was a rabbit anti- IGFBP5 antibody (1:500, #55205-1-AP; Proteintech Group, Chicago, IL, USA).

Techniques: Over Expression, CCK-8 Assay, Migration, Expressing, Control

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Increasing chi-miR-3031 and siRNA-IGFBP5 levels notably decreased IGFBP5 mRNA expression. a : Effect of chi-miR-3031 on IGFBP5 mRNA. b : Effect of siRNA-IGFBP5 on IGFBP5 mRNA. P < 0.01 is shown as **. MC: Chi-miR-3031 mimic. NC: Negative control

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Expressing, Negative Control

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Effects of chi-miR-3031 and siRNA-IGFBP5 on IGFBP5 protein levels. a : Western blot analysis results. b : Densitometric quantification of western blot results. Protein levels were normalized to β-actin. P < 0.05 is shown as *, and P < 0.01 is shown as **. MC: Chi-miR-3031 mimic. NC: Negative control

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Western Blot, Negative Control

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Protein expression levels of κ-casein ( a ) and β-casein ( b ) in GMECs transfected with MC, NC and siRNA-IGFBP5. P < 0.05 is shown as *, and P < 0.01 is shown as **. MC: Chi-miR-3031 mimic. NC: Negative control

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Expressing, Transfection, Negative Control

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Effects of chi-miR-3031 and siRNA-IGFBP5 on p-mTOR protein levels. a : Western blot analysis results. b : Densitometric quantification of western blot results. P < 0.05 is shown as *, and P < 0.01 is shown as **. MC: Chi-miR-3031 mimic. NC: Negative control

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Western Blot, Negative Control

Journal: BMC Veterinary Research

Article Title: Chi-miR-3031 regulates beta-casein via the PI3K/AKT-mTOR signaling pathway in goat mammary epithelial cells (GMECs)

doi: 10.1186/s12917-018-1695-6

Figure Lengend Snippet: Primer information for RT-qPCR

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with a rabbit anti-IGFBP5 polyclonal antibody (1:300, Boster, USA), a rabbit anti-mTOR polyclonal antibody (1:100, Boster, USA), a rabbit anti-p-mTOR polyclonal antibody (1:100, Boster, USA) or a mouse β-actin monoclonal primary antibody (1:1000, Beyotime, Jiangsu, China).

Techniques: Sequencing

Journal: Hepatology (Baltimore, Md.)

Article Title: Association of gankyrin protein expression with early clinical stages and insulin-like growth factor-binding protein 5 expression in human hepatocellular carcinoma.

doi: 10.1002/hep.22027

Figure Lengend Snippet: Fig. 5. Immunohistochemical detection of IGFBP-5 in hepatocellular carcinoma (HCC). HCC sections were stained with anti–IGFBP-5 antibody and counterstained with hematoxylin. Positive immunostaining appears brown. (A) Positive staining for IGFBP-5 in the cytoplasm of HCC cells, especially at the invasive boundaries. (B) Presence of IGFBP-5 in non- cancerous cirrhotic hepatocytes. (C) Stronger staining for IGFBP-5 in HCC cells (upper) than the neighboring cirrhotic hepatocytes (lower). (D) Positive staining for IGFBP-5 in HCC cells (upper left), but negative in cirrhotic cells (lower right). Bar, 100 m. (E) Correlation of expression levels of gankyrin and IGFBP-5 in HCCs. The immunostaining levels were expressed as 0 (negative), 1 (weakly positive), 2 (moderately pos- itive), or 3 (strongly positive). Each diamond represents 1 case. The Spearman’s rho 0.629, P 0.001. (F) Correlation of expression levels of gankyrin and IGFBP-5 in noncancerous hepatocytes determined as in (E). The Spearman’s rho 0.606, P 0.001.

Article Snippet: For immunohistochemistry, mouse monoclonal antigankyrin (3A6C2), anti-MDM2 (Ab-4, Oncogene research products, Boston, MA), and anti-p53 (DO-7, DAKO) antibodies, rabbit polyclonal anti-IGFBP-5 antibody (GroPep, Thebarton, Australia), and horseradish peroxidase–conjugated secondary antibodies against mouse or rabbit immunoglobulins (DAKO) were used.

Techniques: Immunohistochemical staining, Staining, Immunostaining, Expressing

Journal: Hepatology (Baltimore, Md.)

Article Title: Association of gankyrin protein expression with early clinical stages and insulin-like growth factor-binding protein 5 expression in human hepatocellular carcinoma.

doi: 10.1002/hep.22027

Figure Lengend Snippet: Fig. 4. Induction of IGFBP-5 by gankyrin. (A) U-2 OS cells (lanes 1 and 2) and Huh-7 cells (lanes 3 and 4) transiently transfected with plasmids expressing gankyrin or vector alone were analyzed for expres- sion of IGFBP-5 by western blotting using the indicated antibodies. Representative results from more than 3 experiments are shown. (B) Huh-7 cells, mock transfected or transfected with siRNA for gankyrin or control RNA as indicated, were analyzed as in (A). (C) Suppression of IGFBP-5 expression by siRNA. Huh-7 cells were transfected with control RNA or IGFBP-5–specific siRNA. IGFBP-5 transcript levels were deter- mined by real-time RT-PCR and normalized with -actin levels. Results from 3 repeats were averaged and expressed relative to control. Error bars refer to standard deviation of the average quantitated results. (D) Effect of IGFBP-5 down-regulation on cell growth. U-2 OS and Huh-7 cells were transfected with IGFBP-5 siRNA or control RNA, and 72 hours later viable cell numbers were determined. Values are mean standard deviation (n 3) and expressed relative to controls. ** and *, P 0.01 and P 0.05, respectively.

Article Snippet: For immunohistochemistry, mouse monoclonal antigankyrin (3A6C2), anti-MDM2 (Ab-4, Oncogene research products, Boston, MA), and anti-p53 (DO-7, DAKO) antibodies, rabbit polyclonal anti-IGFBP-5 antibody (GroPep, Thebarton, Australia), and horseradish peroxidase–conjugated secondary antibodies against mouse or rabbit immunoglobulins (DAKO) were used.

Techniques: Transfection, Expressing, Plasmid Preparation, Western Blot, Control, Quantitative RT-PCR, Standard Deviation

Journal: Hepatology (Baltimore, Md.)

Article Title: Association of gankyrin protein expression with early clinical stages and insulin-like growth factor-binding protein 5 expression in human hepatocellular carcinoma.

doi: 10.1002/hep.22027

Figure Lengend Snippet: Fig. 6. Survival of patients and expression of molecular markers. The Kaplan-Meier method was used to determine the patient survival and log-rank test to compare survival between patients with HCC grouped according to (A) gankyrin positivity, (B) p53 positivity, (C) MDM2 positivity, and (D) IGFBP-5 positivity. P, positive. N, negative.

Article Snippet: For immunohistochemistry, mouse monoclonal antigankyrin (3A6C2), anti-MDM2 (Ab-4, Oncogene research products, Boston, MA), and anti-p53 (DO-7, DAKO) antibodies, rabbit polyclonal anti-IGFBP-5 antibody (GroPep, Thebarton, Australia), and horseradish peroxidase–conjugated secondary antibodies against mouse or rabbit immunoglobulins (DAKO) were used.

Techniques: Expressing