Journal: American Journal of Physiology - Cell Physiology

Article Title: MiR-4674 regulates angiogenesis in tissue injury by targeting p38K signaling in endothelial cells

doi: 10.1152/ajpcell.00542.2019

Figure Lengend Snippet: Interleukin 1 receptor-associated kinase 1 (IRAK1) and BICD cargo adaptor 2 (BICD2) are bona fide targets of miR-4674 in endothelial cells (ECs). A: discovery and validation of miR-4674 target genes. Human umbilical vein endothelial cells (HUVECs) transfected with miR-negative control (NSm) and miR-4674 mimics (miR-4674m) were subjected to microarray gene profiling. Potential gene targets were further narrowed down by sequential use of bioinformatics and prediction algorithms, RT-quantitative PCR, Western blot analyses, 3′-untranslated region (UTR) reporter studies, and microribonucleoprotein immunoprecipitation (miRNP-IP) analysis. B and C: HUVECs transfected with NSm or miR-4674m were subjected to RT-qPCR for IRAK1 and BICD2 expression (B) or Western blot analyses using antibodies to IRAK1, BICD2, and GAPDH (n = 3 experiments) (C). D: luciferase activity of IRAK1 3′-untranslated region (UTR) and BICD2 3′-UTR normalized to total protein was quantified in HUVECs transfected with NSm, miR-4674m, NSi, or miR-4674i (n = 3 experiments). E: miRNP-IP analysis of enrichment of IRAK1 and BICD2 mRNA in HUVECs transfected with NSm or miR-4674m. *P < 0.01. RT-qPCR was performed to detect IRAK1, BICD2, or SMAD1. Results are representative of n = 3 replicates per group and 2 independent experiments. *P < 0.01. All data represent means ± SE.

Article Snippet: For DNA microarray gene chip analysis, HUVECs were transfected with 30 nM miRNA negative control or miR-4674 mimics for 24 h. Cells were collected into RNeasy mini kit (Qiagen) and sent for two-color, 4 × 44 K format (Agilent Technologies) human whole genome oligo microarray service (ArrayStar).

Techniques: Transfection, Negative Control, Microarray, Real-time Polymerase Chain Reaction, Western Blot, Immunoprecipitation, Quantitative RT-PCR, Expressing, Luciferase, Activity Assay

Journal: Biomolecules

Article Title: Recent Progress in Development and Application of DNA, Protein, Peptide, Glycan, Antibody, and Aptamer Microarrays

doi: 10.3390/biom13040602

Figure Lengend Snippet: Different microarray printing strategies: ( A ) Fabrication of PDA patterned CYTOP (perfluoropolymer) glass slides via photolithography approach (reprinted with permission from ), ( B ) a representative of various steps involved in inkjet printing process (taken from https://gesim-bioinstruments-microfluidics.com/microarray-printer/ ; Accessed on 12 December 2022), and ( C ) GLAD method used to fabricate silver nanorods on glass slides (reprinted with permission from ).

Article Snippet: In various studies, commercially available microarrays, such as the Explorer Antibody Microarray (ASB6000, Full Moon Biosystems, Sunnyvale, CA, USA) with 656 antibodies [ ], VaxArray Coronavirus SeroAssay kit (#VXCV-5100, InDevR, Boulder, CO, USA) [ ], Human Cytokine Antibody Microarray slides (AAH-CYT-G4000 kit, RayBiotech, Peachtree Corners, GA, USA) [ ], Biotin-labeled Human antibody microarray (RayBio Biotech, Guangzhou, China) [ ], Protein Microarray Slides (Grace Bio-Labs, Bend, OR, USA) [ ], HuProt human proteome microarrays (CDI laboratories, Mayaguez, PR, USA) [ ], SBC Mouse ceRNA microarray V1.0 (Shanghai Biotechnology Corporation, Shanghai, China) [ ], and Agilent Whole Mouse 44 K ver.

Techniques: Microarray

Journal: Biomolecules

Article Title: Recent Progress in Development and Application of DNA, Protein, Peptide, Glycan, Antibody, and Aptamer Microarrays

doi: 10.3390/biom13040602

Figure Lengend Snippet: Detection methods: ( A ) FRET based approach to detect target DNA. In presence of target DNA only, the existing complex of fluorescence dye (Cy3) and quencher (BHQ2) is disrupted and generates fluorescence signal (). ( B ) CL based enzymatic approach. This approach involves multiple steps: immunoreaction, HCR amplification, enzyme conjugation, and enzyme-CL (luminol p-iodophenol) substrate reaction (). ( C ) Colorimetric detection (use of streptavidin-coated gold nanoparticles) of isothermal DNA amplification. Interaction of biotin and streptavidin was explored (). ( D ) OIRD, a label-free detection method to study protein/antibody interactions with target molecules on a protein microarray chip ().

Article Snippet: In various studies, commercially available microarrays, such as the Explorer Antibody Microarray (ASB6000, Full Moon Biosystems, Sunnyvale, CA, USA) with 656 antibodies [ ], VaxArray Coronavirus SeroAssay kit (#VXCV-5100, InDevR, Boulder, CO, USA) [ ], Human Cytokine Antibody Microarray slides (AAH-CYT-G4000 kit, RayBiotech, Peachtree Corners, GA, USA) [ ], Biotin-labeled Human antibody microarray (RayBio Biotech, Guangzhou, China) [ ], Protein Microarray Slides (Grace Bio-Labs, Bend, OR, USA) [ ], HuProt human proteome microarrays (CDI laboratories, Mayaguez, PR, USA) [ ], SBC Mouse ceRNA microarray V1.0 (Shanghai Biotechnology Corporation, Shanghai, China) [ ], and Agilent Whole Mouse 44 K ver.

Techniques: Fluorescence, Amplification, Conjugation Assay, DNA Amplification, Microarray

Journal: Biomolecules

Article Title: Recent Progress in Development and Application of DNA, Protein, Peptide, Glycan, Antibody, and Aptamer Microarrays

doi: 10.3390/biom13040602

Figure Lengend Snippet: ( A ) A pictorial workflow presentation of hybrid tetrahedral DNA structured probe in conjugation with hybridization chain reaction (DTSP-HCR) concept used to distinguish single-base mismatches in DNA () and ( B ) an overview of acute tuberculosis (ATB) biomarker identification using a two-phase strategy (discovery phase and validation phase) (). Venn diagram and microarray chip visual analysis revealed the potential of 5 protein biomarkers to distinguish ATB and LTBI/HC. LTBI represents latent tuberculosis infection, and HC represents healthy control.

Article Snippet: In various studies, commercially available microarrays, such as the Explorer Antibody Microarray (ASB6000, Full Moon Biosystems, Sunnyvale, CA, USA) with 656 antibodies [ ], VaxArray Coronavirus SeroAssay kit (#VXCV-5100, InDevR, Boulder, CO, USA) [ ], Human Cytokine Antibody Microarray slides (AAH-CYT-G4000 kit, RayBiotech, Peachtree Corners, GA, USA) [ ], Biotin-labeled Human antibody microarray (RayBio Biotech, Guangzhou, China) [ ], Protein Microarray Slides (Grace Bio-Labs, Bend, OR, USA) [ ], HuProt human proteome microarrays (CDI laboratories, Mayaguez, PR, USA) [ ], SBC Mouse ceRNA microarray V1.0 (Shanghai Biotechnology Corporation, Shanghai, China) [ ], and Agilent Whole Mouse 44 K ver.

Techniques: Conjugation Assay, Hybridization, Biomarker Discovery, Microarray, Infection, Control

Journal: Biomolecules

Article Title: Recent Progress in Development and Application of DNA, Protein, Peptide, Glycan, Antibody, and Aptamer Microarrays

doi: 10.3390/biom13040602

Figure Lengend Snippet: A generic workflow of the application of glycan microarray chip to efficiently profile enzyme activities and determine enzyme inhibitor IC 50 values ().

Article Snippet: In various studies, commercially available microarrays, such as the Explorer Antibody Microarray (ASB6000, Full Moon Biosystems, Sunnyvale, CA, USA) with 656 antibodies [ ], VaxArray Coronavirus SeroAssay kit (#VXCV-5100, InDevR, Boulder, CO, USA) [ ], Human Cytokine Antibody Microarray slides (AAH-CYT-G4000 kit, RayBiotech, Peachtree Corners, GA, USA) [ ], Biotin-labeled Human antibody microarray (RayBio Biotech, Guangzhou, China) [ ], Protein Microarray Slides (Grace Bio-Labs, Bend, OR, USA) [ ], HuProt human proteome microarrays (CDI laboratories, Mayaguez, PR, USA) [ ], SBC Mouse ceRNA microarray V1.0 (Shanghai Biotechnology Corporation, Shanghai, China) [ ], and Agilent Whole Mouse 44 K ver.

Techniques: Glycoproteomics, Microarray