gelatin coated ecis arrays  (Applied BioPhysics)

Journal: Journal of Cell Communication and Signaling

Article Title: Critical role of mitogen-inducible gene 6 in restraining endothelial cell permeability to maintain vascular homeostasis

doi: 10.1007/s12079-022-00704-z

Figure Lengend Snippet: Vascular permeability is increased in Mig6 knockout mice and endothelial barrier is compromised by MIG6 knockdown. ( a ) Representative images of WT (top) and Mig6 −/− (bottom) mouse showing Evans blue leakage, after treatment with 50 ng/ml of VEGFA (left) or BSA (right) for 1 h. ( b ) Images of Evans blue leakage in excised ears of WT and Mig6 −/− mouse. ( c ) Quantification of Evans blue leakage in ear tissues. Evans blue dye was extracted from ear skin of WT and Mig6 −/− mice to read an absorbance ( n = 3 – 5). ( d ) Histamine (100 μM) or VEGFA (50 ng/ml)-induced EC permeability in siControl or siMIG6-treated HUVECs ( n = 4). ( e ) Normalized resistance of siControl or siMIG6-treated ECs after stimulation with VEGFA. HUVECs were treated with siControl or siMIG6, and grown to confluence on gelatin-coated electrode arrays. After low serum starvation for 3 h, EC monolayer was stimulated with VEGFA (50 ng/ml) and TEER was measured by ECIS at a frequency of 4000 Hz. One-way ANOVA with Sidak multiple comparison test ( c , d) and unpaired Student’s t-test ( e ) were performed. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, NS: not statistically significant

Article Snippet: MIG6 knockdown ECs were seeded onto gelatin-coated 8W10E ECIS array (Applied Biophysics, MA, USA) and allowed to reach confluence overnight.

Techniques: Permeability, Knock-Out

Journal: The FASEB Journal

Article Title: Resident mesenchymal vascular progenitors modulate adaptive angiogenesis and pulmonary remodeling via regulation of canonical Wnt signaling

doi: 10.1096/fj.202000629R

Figure Lengend Snippet: Disrupted Wnt signaling in ABCG2 pos COPD MVPC disrupts MVEC function. A, Heatmap for differentially expressed genes in MVPC from COPD vs non‐diseased lungs (color scale shown at the top right) B and C, Validation of representative genes expressed in COPD ABCG2 pos lung MVPC, compared to control, enriched in GO categories of B, Wnt signaling and C, actin binding, contractility, and migration. The mean of combined patient samples per group as well as results for individual samples are presented. n = 3‐6 patient samples per group. D, Western blot analysis of Dkk1, Wnt5a and Slit2 protein expression. n = 3, 3. Data presented as the mean ± standard error of mean (SEM). E, Cocultures of pulmonary MVEC and MVPC were analyzed for the effect of control or COPD MVPC on barrier function following injury. MVPC were plated on a monolayer of MVEC at a ratio of 3:1. The groups underwent no injury or an electrical wounding injury using the ECIS system. The presence of COPD MVPC following injury decreased the rate at which barrier formation is recovered. Quantitation of normalized resistance at indicated time points (Δ) was presented in bar graph format. Data presented as mean (±SEM). Controls included MVEC alone and uninjured MVEC. n = 4. F, Murine WT and Wnt activated βOE MVPC were analyzed by PCR to examine the expression of angiogenic transcripts identified as different between control and COPD samples. n = 3, 3. All amplification was normalized to a housekeeping gene and the results presented as mean fold change over control. Data presented as mean ± SEM. G. Representative summary of murine and human PCR data

Article Snippet: Human lung MVECs (Lonza, Walkersville) were plated at a concentration of 112 500 cells per well on gelatin coated 8W1E PET ECIS culture ware arrays (Applied Biophysics, Troy) overnight to achieve confluence.

Techniques: Biomarker Discovery, Control, Binding Assay, Migration, Western Blot, Expressing, Quantitation Assay, Amplification