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: FIGURE 5 Disrupted Wnt signaling in ABCG2pos 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 ABCG2pos 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

Journal: The Journal of biological chemistry

Article Title: An siRNA library screen identifies CYLD and USP34 as deubiquitinases that regulate GPCR-p38 MAPK signaling and distinct inflammatory responses.

doi: 10.1016/j.jbc.2023.105370

Figure Lengend Snippet: Figure 7. Depletion of CYLD but not USP34 enhances thrombin-induced endothelial barrier permeability. Endothelial EA.hy926 cells transfected with nonspecific (ns), CYLD- (A), and USP34- (B) specific siRNA were stimulated with or without thrombin (Th), and endothelial barrier permeability (resistance) was measured by ECIS. Changes in endothelial barrier resistance were normalized to baseline for each condition, and data expressed relative to maximum thrombin-induced change in resistance quantified from three independent experiments for CYLD and USP34 siRNA knockdown. The line graphs are from a representative experiment (mean ±S.D., performed in triplicate, n=3) for each DUB knockdown. The bar graphs are the peak change in resistance (mean ± S.D.) from the three independent experiments (n=3) and were analyzed by two-tailed student’s t test. P-value: a=0.0415; ns, not significant. C, cell lysates were prepared from the nonspecific, CYLD and USP34 siRNA transfected cells and immunoblotted as indicated. D, cartoon illustration of thrombin- activated PAR1 ubiquitin-driven p38 mediated endothelial barrier disruption regulation by CYLD.

Article Snippet: Electrical cell impedance sensing Endothelial EA.hy926 cells were seeded onto cysteine- and collagen-coated gold microwell 8W10E+ ECIS array (Applied Biophysics).

Techniques: Permeability, Transfection, Knockdown, Two Tailed Test, Ubiquitin Proteomics, Disruption

Journal: International Journal of Molecular Sciences

Article Title: Trefoil Factor 3 (TFF3) Is Involved in Cell Migration for Skeletal Repair

doi: 10.3390/ijms20174277

Figure Lengend Snippet: Influence of TFF3 on cell migration. Electric cell-substrate impedance sensing (ECIS) analyzed the 2D-cell migration over a 90 h period following electrical wounding or in non-wounded controls in the presence or absence of rhTFF3. A representative time-impedance diagram with the relevant time points for statistical analysis is shown in ( a ). Increased cell migration into wounded areas correlated inversely with electrical resistance ( b ). Radius migration assay confirms rapid and even ingrowth of cells stimulated by TFF3 ( c ). Bars show the mean ± SD. * p < 0.05.

Article Snippet: Wound healing was analyzed by the electric cell-substrate impedance sensing (ECIS) instrument Z theta+16 (Applied Biophysics, New York, USA) using ECIS culture ware 8W2LE arrays.

Techniques: Migration, Electric Cell-substrate Impedance Sensing