Journal: Cells

Article Title: miR-23a-3p is a Key Regulator of IL-17C-Induced Tumor Angiogenesis in Colorectal Cancer

doi: 10.3390/cells9061363

Figure Lengend Snippet: IL-17C stimulation promotes human intestinal microvascular endothelial cells (HIMEC) angiogenesis. ( A ) Representative bands of RT-PCR and quantitative analyses of two receptors of IL-17C, IL-17RA ( A , left ), and IL-17RE ( A , right ) in HIMECs treated with vehicle control or IL-17C (100 ng/mL, 3 h) are shown ( n = 3). GAPDH was used for the loading control. NS, not significant. ( B ) HIMECs were seeded into polymerized Matrigel and treated with IL-17C (100 ng/mL, 4 h) for in vitro tube formation assays ( B , left ). Scale bar, 100 μm. Total tube length of photos was measured using ImageJ v.1.47 software ( n = 6) ( B , right ). Error bars, SD. ** p < 0.01 vs. the control group. ( C ) The mouse aortic ring assay was conducted to investigate ex vivo angiogenic activity of IL-17C (100 ng/mL, 14 d) ( C , top ). Scale bar, 50 μm. For quantification, total sprouted tube vessels of 3 independent photos from each group were measured using ImageJ v.1.47 software ( C , bottom ). Error bars, SEM ( n = 3). *** p < 0.001 vs. the control group. ( D ) For the wound healing assays, cells were seeded in a culture plate, incubated with culture media with/without IL-17C (100 ng/mL), and scratched to create wounds. Photos were taken 24 h after wound creation ( D , left ). Scale bar, 50 μm. A view of representative fields from images of the wound healing assays, analyzed using ImageJ v.1.47 software ( D , right ). Error bars, SD ( n = 6). ** p < 0.01 vs. the control group. ( E ) F-actin organization in endothelial cells determined via rhodamine-phalloidin staining ( n = 3). Scale bar, 50 μm. ( F ) PCR microarray was performed to investigate expression levels of angiogenesis-related genes by IL-17C (100 ng/mL, 3 h) treatment in HIMECs ( n = 1). ( G ) The mRNA levels of CXCL3, IL-8, MMP-1, and MMP-10 were analyzed by qPCR at 3 h post IL-17C treatment. Error bars, SD ( n = 3). ** p < 0.01 and *** p < 0.001 vs. the control group.

Article Snippet: Briefly, the cells were then washed with PBS, fixed with 10% formalin (Sigma-Aldrich) for 15 min, and permeabilized with Triton X-100 (0.1%, Daejung, Gyeonggi-do, South Korea) for 5 min. After permeabilization, the cells were washed twice and stained for 15 min in the dark with rhodamine–phalloidin (100 nM/well, Cytoskeleton, Denver, CO, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, In Vitro, Software, Aortic Ring Assay, Ex Vivo, Activity Assay, Incubation, Staining, Microarray, Expressing

Journal: Scientific Reports

Article Title: Magnetically-driven 2D cells organization on superparamagnetic micromagnets fabricated by laser direct writing

doi: 10.1038/s41598-020-73414-4

Figure Lengend Snippet: Scanning electron micrographs of 2D microarray fabricated by LDW via TPP: ( a ) Ormo squares; ( b ) Ormo/MNPs squares; ( c ) Ormo superposed with Ormo/MNPs squares forming the chessboard-like 2D microarray; ( d ) inset from ( a ); ( e ) inset from ( b ); ( f ) inset from ( c ) showing Ormo/MNPs square (left) and Ormo square (right); ( g ) Inset from ( c ) showing the intersection of 4 squares from the 2D microarray; DHM images of 2D microarray: ( h ) 3D reconstructed hologram; ( i ) phase shift 2D map at the intersection of 4 squares; ( j ) line profile of the height difference between adjacent Ormo/MNPs and Ormo squares along the green line from ( i ).

Article Snippet: Figure 2 Images of 2D microarray obtained by enhanced dark field microscopy using the Cytoviva 3D module (MNPs represented as yellow dots (false colors)): ( a ) top view; ( b ) tilted view; ( c – e ) EDX mapping of iron, carbon and oxygen.

Techniques: Microarray

Journal: Scientific Reports

Article Title: Magnetically-driven 2D cells organization on superparamagnetic micromagnets fabricated by laser direct writing

doi: 10.1038/s41598-020-73414-4

Figure Lengend Snippet: Images of 2D microarray obtained by enhanced dark field microscopy using the Cytoviva 3D module (MNPs represented as yellow dots (false colors)): ( a ) top view; ( b ) tilted view; ( c – e ) EDX mapping of iron, carbon and oxygen.

Article Snippet: Figure 2 Images of 2D microarray obtained by enhanced dark field microscopy using the Cytoviva 3D module (MNPs represented as yellow dots (false colors)): ( a ) top view; ( b ) tilted view; ( c – e ) EDX mapping of iron, carbon and oxygen.

Techniques: Microarray, Microscopy

Journal: Scientific Reports

Article Title: Magnetically-driven 2D cells organization on superparamagnetic micromagnets fabricated by laser direct writing

doi: 10.1038/s41598-020-73414-4

Figure Lengend Snippet: 2D microarrays: ( a , f ) Optical images; ( b , g ) Green autofluorescence; Representative fluorescence microscope images of fibroblasts seeded on 2D microarrays: ( c , h ) cytoskeleton (red) visualized by staining with F-actin; ( d , i ) nuclei (blue) visualized by staining with Hoechst; ( e , j ) overlapped fluorescence images of 2D microarrays, cells cytoskeleton and nuclei; ( k , m ) 3D top and ( l , n ) 3D tilted views of cells nuclei (blue), cytoskeleton (red) and centers of MNPs and agglomerations of MNPs (yellow), imaged by enhanced dark field microscopy. The samples were imaged in the presence ( a – e , k , l ) and absence ( f – j , m , n ) of SMF. All cells were imaged after 1 day of cell culture.

Article Snippet: Figure 2 Images of 2D microarray obtained by enhanced dark field microscopy using the Cytoviva 3D module (MNPs represented as yellow dots (false colors)): ( a ) top view; ( b ) tilted view; ( c – e ) EDX mapping of iron, carbon and oxygen.

Techniques: Fluorescence, Microscopy, Staining, Cell Culture

Journal: Developmental cell

Article Title: YAP/TAZ and Hedgehog coordinate growth and patterning in gastrointestinal mesenchyme

doi: 10.1016/j.devcel.2017.08.019

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: HRP conjugated donkey anti-rabbit , Jackson Immunoresearch , 711-035-152.

Techniques: Recombinant, Plasmid Preparation, Reporter Assay, Reverse Transcription, Microarray, Sequencing, Software

Journal: Life science alliance

Article Title: Inflammatory risk contributes to post-COVID endothelial dysfunction through anti-ACKR1 autoantibody.

doi: 10.26508/lsa.202402598

Figure Lengend Snippet: Figure 1. Tissue and cell type expressions of endothelial antigens. (A) Gene expressions of endothelial antigens upon TNFα (10 ng/ml) treatment on human umbilical artery and vein cells for 24 h. Data represent mean ± s.d.; n = 3 biological replicates; two-way ANOVA; *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. (B) Flow cytometry analysis of ACKR1 protein expression on artery and vein cells after 48 h of stimulation with TNFα (10 ng/ml). Data represent mean ± s.d.; n = 3 biological replicates; two-way ANOVA; ****P < 0.0001. (C) Expression profile of Ackr1 in vascular endothelial subtypes of various organs from a mouse single-cell transcriptome atlas (accession code: E-MTAB-8077). Data were generated with EC Atlas web- based visualization.

Article Snippet: Protein microarray for autoantibody detection We custom made a microarray-based autoantibody detection kit (RayBiotech) where a glass slide surface was spotted with purified human recombinant ACKR1 protein (Cat #TP304680; OriGene) that could be recognized by ACKR1 autoantibodies present in human plasma.

Techniques: Flow Cytometry, Expressing, Generated

Journal: Life science alliance

Article Title: Inflammatory risk contributes to post-COVID endothelial dysfunction through anti-ACKR1 autoantibody.

doi: 10.26508/lsa.202402598

Figure Lengend Snippet: Figure 3. Anti-ACKR1 autoantibodies induce endothelial dysfunctions. (A) Experimental workflow of intervening autoantibody-antigen interactions to investigate the functional impact of anti-ACKR1 autoantibodies on human endothelial cells. The three-dimensional structure of ACKR1 was produced using I-TASSER server. (B) Flow cytometry analysis of the percentage of necrotic and late apoptotic endothelial cells after 24 h of experimental treatments. (C) Transendothelial electrical resistance assay to evaluate endothelial barrier permeability after experimental treatments. (D) Number of transmigrated PBMCs in an endothelial-immune cell co-culture transwell assay. (E) Degree of cytotoxicity was determined by quantifying lactate dehydrogenase activity in cell-free supernatants obtained from endothelial cells cultured with purified immunoglobulin G (IgG) and/or PBMCs. (F) Degree of antibody-dependent cellular cytotoxicity was determined from endothelial cells exposed to purified IgG and PBMCs. Purifie IgG was pre-incubated with and without blocking peptide or liposome ACKR1. For (B, C, D, E, F), data represent mean ± s.d.; one-way ANOVA; *P < 0.05, **P < 0.01; ***P < 0.001, ****P < 0.0001; ns, non-significant. Data points represent biological replicates.

Article Snippet: Protein microarray for autoantibody detection We custom made a microarray-based autoantibody detection kit (RayBiotech) where a glass slide surface was spotted with purified human recombinant ACKR1 protein (Cat #TP304680; OriGene) that could be recognized by ACKR1 autoantibodies present in human plasma.

Techniques: Functional Assay, Produced, Flow Cytometry, Permeability, Co-Culture Assay, Transwell Assay, Activity Assay, Cell Culture, Incubation, Blocking Assay