Journal: BioMed Research International

Article Title: Establishment and Verification of a Gene Signature for Diagnosing Type 2 Diabetics by WGCNA, LASSO Analysis, and In Vitro Experiments

doi: 10.1155/2022/4446342

Figure Lengend Snippet: Correlation between the genes in the LASSO model and immune cell infiltrations in type 2 diabetic islets. (a) Heatmap for the associations between hub genes and immune cell infiltrations. Red: positive correlation and blue: negative correlation. (b) Associations between BST2 expression and immune cell infiltrations. (c) Associations between BTBD1 expression and immune cell infiltrations. (d) Associations between IFIT1 expression and immune cell infiltrations. (e) Associations between IFIT3 expression and immune cell infiltrations. (f) Associations between RTP4 expression and immune cell infiltrations. The bigger the circle, the stronger the correlation.

Article Snippet: Two hours later, the membrane was incubated by primary antibodies against BST2 (1/1000; #19277; Cell Signaling Technology, USA), BTBD1 (1/1000; #ab138507; Abcam, USA), TGF- β (1/1000; #84912; Cell Signaling Technology, USA), P53 (1/1000; #2527; Cell Signaling Technology, USA), and GAPDH (1/1000; #5174; Cell Signaling Technology, USA) at 4°C overnight, followed by being incubated by secondary antibodies (1/5 000; ab709; Abcam, USA).

Techniques: Expressing

Journal: BioMed Research International

Article Title: Establishment and Verification of a Gene Signature for Diagnosing Type 2 Diabetics by WGCNA, LASSO Analysis, and In Vitro Experiments

doi: 10.1155/2022/4446342

Figure Lengend Snippet: Expression and involved signaling pathways for the genes in the LASSO model in type 2 diabetic islets. (a) Box plot of the expression of the genes in the LASSO model BST2, BTBD1, IFIT1, IFIT3, and RTP4 between type 2 diabetic islets and normal islets. Ns: Not significant; ∗∗ p < 0.01. (b)–(f) GSEA for identifying the signaling pathways that were positively associated with (b) BST2, (c) BTBD1, (d) IFIT1, (e) IFIT3, and (f) RTP4.

Article Snippet: Two hours later, the membrane was incubated by primary antibodies against BST2 (1/1000; #19277; Cell Signaling Technology, USA), BTBD1 (1/1000; #ab138507; Abcam, USA), TGF- β (1/1000; #84912; Cell Signaling Technology, USA), P53 (1/1000; #2527; Cell Signaling Technology, USA), and GAPDH (1/1000; #5174; Cell Signaling Technology, USA) at 4°C overnight, followed by being incubated by secondary antibodies (1/5 000; ab709; Abcam, USA).

Techniques: Expressing

Journal: BioMed Research International

Article Title: Establishment and Verification of a Gene Signature for Diagnosing Type 2 Diabetics by WGCNA, LASSO Analysis, and In Vitro Experiments

doi: 10.1155/2022/4446342

Figure Lengend Snippet: Validation of the expression of the genes in the LASSO model in glucotoxicity models and normal islet β cells. (a) and (b) RT-qPCR for validating the mRNA expression of (a) BST2 and (b) BTBD1 in glucotoxicity models and normal islet β cells. (c) Representative images of western blot of BST2 and BTBD1 in glucotoxicity models and normal islet β cells. (d) and (e) Quantification results of BST2 and BTBD1 expression in two groups according to western blot. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: Two hours later, the membrane was incubated by primary antibodies against BST2 (1/1000; #19277; Cell Signaling Technology, USA), BTBD1 (1/1000; #ab138507; Abcam, USA), TGF- β (1/1000; #84912; Cell Signaling Technology, USA), P53 (1/1000; #2527; Cell Signaling Technology, USA), and GAPDH (1/1000; #5174; Cell Signaling Technology, USA) at 4°C overnight, followed by being incubated by secondary antibodies (1/5 000; ab709; Abcam, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: BioMed Research International

Article Title: Establishment and Verification of a Gene Signature for Diagnosing Type 2 Diabetics by WGCNA, LASSO Analysis, and In Vitro Experiments

doi: 10.1155/2022/4446342

Figure Lengend Snippet: Silencing BST2 ameliorates apoptosis of glucotoxicity islet β cell models and influences the activation of TGF- β and P53 pathways. (a) RT-qPCR of the expression of BST2 mRNA in islet β cells with si-NC or si-BST2 transfection. (b) and (c) Flow cytometry of apoptotic levels of glucotoxicity models and normal islet β cells. (d) and (e) Flow cytometry of apoptotic levels of glucotoxicity models under si-NC or si-BST2 transfection. (f)–(h) Western blot of the expression of TGF- β and P53 proteins in glucotoxicity models and normal islet β cells. (i)–(k) Western blot of the expression of TGF- β and P53 proteins in glucotoxicity models under si-NC or si-BST2 transfection. ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: Two hours later, the membrane was incubated by primary antibodies against BST2 (1/1000; #19277; Cell Signaling Technology, USA), BTBD1 (1/1000; #ab138507; Abcam, USA), TGF- β (1/1000; #84912; Cell Signaling Technology, USA), P53 (1/1000; #2527; Cell Signaling Technology, USA), and GAPDH (1/1000; #5174; Cell Signaling Technology, USA) at 4°C overnight, followed by being incubated by secondary antibodies (1/5 000; ab709; Abcam, USA).

Techniques: Activation Assay, Quantitative RT-PCR, Expressing, Transfection, Flow Cytometry, Western Blot

Journal: Journal of Immunology Research

Article Title: Activation of FGD5-AS1 Promotes Progression of Cervical Cancer through Regulating BST2 to Inhibit Macrophage M1 Polarization

doi: 10.1155/2021/5857214

Figure Lengend Snippet: Primers for real-time PCR.

Article Snippet: Antibodies against human BST2 and GAPDH were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques:

Journal: Journal of Immunology Research

Article Title: Activation of FGD5-AS1 Promotes Progression of Cervical Cancer through Regulating BST2 to Inhibit Macrophage M1 Polarization

doi: 10.1155/2021/5857214

Figure Lengend Snippet: FGD5-AS1 and BST2 expression in cervical cancer cells. (a) RNA levels of FGD5-AS1 in HeLa, SiHa, C33A, CasKi, and H8 cells. (b) RNA levels of BST2 in HeLa, SiHa, C33A, CasKi, and H8 cells. ∗∗∗ P < 0.001 and ∗ P < 0.05. Three independent assays were carried out.

Article Snippet: Antibodies against human BST2 and GAPDH were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques: Expressing

Journal: Journal of Immunology Research

Article Title: Activation of FGD5-AS1 Promotes Progression of Cervical Cancer through Regulating BST2 to Inhibit Macrophage M1 Polarization

doi: 10.1155/2021/5857214

Figure Lengend Snippet: Effects of FGD5-AS1 on cervical cancer cell aggressiveness. (a) FGD5-AS1 expression in SiHa cells. FGD5-AS1 knockdown was generated in the SiHa cells by transfection of shFGD5-AS1. The transfection efficiency was validated by real-time PCR. (b, c) BST2 mRNA and protein expression in SiHa cells. (d) Cell viability by CCK8 assay. (e) Cell apoptosis by TUNEL assay. (f) Migration and invasion capacity using Transwell assay. ∗ P < 0.05. shNC shRNA: negative control. Three independent assays were carried out.

Article Snippet: Antibodies against human BST2 and GAPDH were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques: Expressing, Generated, Transfection, Real-time Polymerase Chain Reaction, CCK-8 Assay, TUNEL Assay, Migration, Transwell Assay, shRNA, Negative Control

Journal: Journal of Immunology Research

Article Title: Activation of FGD5-AS1 Promotes Progression of Cervical Cancer through Regulating BST2 to Inhibit Macrophage M1 Polarization

doi: 10.1155/2021/5857214

Figure Lengend Snippet: Effects of FGD5-AS1 on M2-like polarization of macrophages. (a) The expression of FGD5-AS1 was increased in M2 macrophages. (b) The expression of BST2 was upregulated in M2 macrophages. M0 cells were transfected with FGD5-AS1 shRNA. (c) Expression of M1 markers (CD80 and CD86). (d) Expression of M2 markers (CD206 and CD163). (e) Flow cytometry analysis of CD80 and CD86 in different transfectants of M0 cells. (f) Flow cytometry analysis of CD206 and CD163 in different transfectants of M0 cells. ∗ P < 0.05. Three independent assays were carried out.

Article Snippet: Antibodies against human BST2 and GAPDH were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques: Expressing, Transfection, shRNA, Flow Cytometry

Journal: Journal of Immunology Research

Article Title: Activation of FGD5-AS1 Promotes Progression of Cervical Cancer through Regulating BST2 to Inhibit Macrophage M1 Polarization

doi: 10.1155/2021/5857214

Figure Lengend Snippet: FGD5-AS1 sponged miR-129-5p to regulate BST2. (a) Binding site between FGD5-AS1 and miR-129-5p. (b) The luciferase activity in FGD5-AS1-WT and FGD5-AS1-MUT after transfection with miR-129-5p mimics or NC. (c, d) Enrichment of FGD5-AS1 and miR-129-5p in the anti-Ago2 or IgG immunoprecipitates in SiHa cells transfected with miR-129-5p NC or inhibitors. (e) The predicted binding site between BST2 and miR-129-5p. (f) The luciferase activity in BST2-WT and BST2-MUT after transfection with miR-129-5p mimics or NC. ∗ P < 0.05. Three independent assays were carried out.

Article Snippet: Antibodies against human BST2 and GAPDH were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection

Journal: Journal of Immunology Research

Article Title: Activation of FGD5-AS1 Promotes Progression of Cervical Cancer through Regulating BST2 to Inhibit Macrophage M1 Polarization

doi: 10.1155/2021/5857214

Figure Lengend Snippet: BST2 promoted cervical cancer progression via inducing M2 macrophage polarization. (a) mRNA levels of BST2 in SiHa cells transfected with BST2 shRNA. (b) Cell viability by CCK8 assay. (c) Cell apoptosis by TUNEL assay. (d) Migration and invasion capacity by Transwell assay. (e) mRNA expression of CD80 and CD86 in macrophages. (f) mRNA expression of CD206 and CD163 in M0 macrophages transfected with BST2 shRNA. ∗ P < 0.05. Three independent assays were carried out.

Article Snippet: Antibodies against human BST2 and GAPDH were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques: Transfection, shRNA, CCK-8 Assay, TUNEL Assay, Migration, Transwell Assay, Expressing

Journal: Journal of Immunology Research

Article Title: Activation of FGD5-AS1 Promotes Progression of Cervical Cancer through Regulating BST2 to Inhibit Macrophage M1 Polarization

doi: 10.1155/2021/5857214

Figure Lengend Snippet: miR-129-5p mimics reversed FGD5-AS1-induced upregulation of BST2 and M2 macrophage polarization. (a) Levels of FGD5-AS1 in SiHa cells transfected with vector, FGD5-AS1, FGD5-AS1 plus miR-129-5p NC, or FGD5-AS1 plus miR-129-5p mimics. (b, c) The levels of BST2 in SiHa cells. (d) mRNA expression of CD80 and CD86 in macrophages. (e) mRNA expression of CD206 and CD163 in macrophages. ∗ P < 0.05. Three independent assays were carried out.

Article Snippet: Antibodies against human BST2 and GAPDH were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques: Transfection, Plasmid Preparation, Expressing