Journal:

Article Title: The Mitochondrial Toxin Produced by Streptomyces griseus Strains Isolated from an Indoor Environment Is Valinomycin

doi:

Figure Lengend Snippet: Toxicity to boar spermatozoa of methanol-soluble substances from actinomycetes isolated from indoor environments

Article Snippet: Twelve strains were identified by chemotaxonomic methods and by 16S rDNA sequencing as members of the genera Streptomyces , Nocardiopsis , and Dietzia (Table ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain Toxicity to boar spermatozoa a Source of strain b EC 50 of methanol-soluble substances (ng ml −1 ) EC 50 of cell extract (mg ml −1 ) Indoor isolates S. griseus 2/ppi 20 0.0005 (10 5 ) c Children’s day care center (settled dust) S. griseus 8/ppi 50 0.001 (10 6 ) Children’s day care center (settled dust) S. griseus 10/ppi 10 0.0009 (10 6 ) Children’s day care center (settled dust) S. griseus 1/k 63 0.002 (10 6 ) Elementary school (air) Nocardiopsis albus 123 7,500 0.5 (10 8 ) Children’s day care center (water-damaged building material) S. griseus 157 59,000 >9 (>10 9 ) Children’s day care center (water-damaged building material) Dietzia sp. strain 147 >72,000 >1 (>10 9 ) Children’s day care center (water-damaged building material) Dietzia sp. strain 148 >86,000 >1 (>10 9 ) Children’s day care center (water-damaged building material) Nocardiopsis dassonvillei 305 >10,000 >0.3 (>10 8 ) Cattle barn (air) Streptomyces albidoflavus 703 36,000 1 (10 8 ) Cattle barn (settled dust) Nocardiopsis dassonvillei 704 >40,000 >0.5 (>10 8 ) Cattle barn (air) Reference strains S. griseus DSM 40236 T 26,000 1 (10 8 ) DSMZ Dietzia maris N 1009 T <36,000 >1 (>10 8 ) DSMZ Bacillus mycoides ATCC 6462 T 67,000 2.5 (10 8 ) ATCC B. cereus ATCC 14569 T 16,000 1 (10 8 ) ATCC B. cereus 4810/72 c 28 0.002 (10 6 ) Food poisoning (emetic) Open in a separate window a The EC 50 is the concentration that paralyzed more than 50% of the sperm cells. b DSMZ, Deutsche Sammlung von Mikroorganism und Zellkulturen GmbH; ATCC, American Type Culture Collection. c The numbers in parentheses are the equivalent numbers of cells. d Data from reference 3 .

Techniques: Isolation

Journal: APL Bioengineering

Article Title: Cell shape, and not 2D migration, predicts extracellular matrix-driven 3D cell invasion in breast cancer

doi: 10.1063/1.5143779

Figure Lengend Snippet: ECM proteins upregulated in breast tumor tissue have distinct cell line-specific effects on tumor cell adhesion. (a) Representative images of MDA-MB-231 cells plated on plastic, Collagen I, Fibronectin, Tenascin C, or Collagen IV for 2 h, fixed and stained with Phalloidin (red) and DAPI (blue). The scale bar is 100 μ m. Quantification of cell shape features using Cell Profiler to evaluate effects on MDA-MB-231: (b) area/cell (10 3 μ m 2 ), (c) eccentricity, and (d) compactness. Results show entire distribution, no ECM (n = 793 cells), Collagen I (n = 907 cells), Fibronectin (n = 1154 cells), Tenascin C (n = 104 cells), or Collagen IV (n = 766 cells). Significance by one-way ANOVA, *** p < 0.005. (e) Representative images of MDA-MB-468 cells plated on plastic, Collagen I, Fibronectin, Tenascin C, or Collagen IV for 2 h, fixed and stained with Phalloidin (red) and DAPI (blue). Scale bar is 100 μ m. Quantification of cell shape features using Cell Profiler to evaluate effects on MDA-MB-468: (f) area/cell (10 3 μ m 2 ), (g) eccentricity, and (h) compactness. Results show entire distribution, no ECM (n = 802 cells), Collagen I (n = 1016 cells), Fibronectin (n = 945 cells), Tenascin C (n = 101 cells), or Collagen IV (n = 1073 cells). Significance by one-way ANOVA, *** p < 0.005, ns is not significant.

Article Snippet: We used the following ECM proteins: Collagen I protein (CB-40236; Fisher Scientific, Hampton, NH), Fibronectin protein (F1141; SIGMA, St. Louis, MO), Tenascin C (R&D systems, 3358TC050), Collagen IV protein (Abcam, ab7536), and Matrigel (growth-factor reduced, Corning, CB-40230C).

Techniques: Staining

Journal: APL Bioengineering

Article Title: Cell shape, and not 2D migration, predicts extracellular matrix-driven 3D cell invasion in breast cancer

doi: 10.1063/1.5143779

Figure Lengend Snippet: Clustering of adhesion parameters reveals ECM-specific effects on cell shape. (a) Visualization of continuum of cell adhesion of MDA-MB-231 cells on different ECM substrates based on all 11 cell shape parameters with SPRING plots. (b) Plots showing localization of the ECM factor-dependent cell adhesion shape on the combined SPRING plot. Mean centered cell adhesion of MDA-MB-231 (c) and MDA-MB-468 (d) cells plated on plastic, Collagen I, Fibronectin, Tenascin C, or Collagen IV for 2 h. Each cell adhesion parameter and ECM factor are clustered by rank correlation and mean linkage.

Article Snippet: We used the following ECM proteins: Collagen I protein (CB-40236; Fisher Scientific, Hampton, NH), Fibronectin protein (F1141; SIGMA, St. Louis, MO), Tenascin C (R&D systems, 3358TC050), Collagen IV protein (Abcam, ab7536), and Matrigel (growth-factor reduced, Corning, CB-40230C).

Techniques:

Journal: APL Bioengineering

Article Title: Cell shape, and not 2D migration, predicts extracellular matrix-driven 3D cell invasion in breast cancer

doi: 10.1063/1.5143779

Figure Lengend Snippet: ECM-driven effects on 2D cell migration speed do not correlate with effects on persistence. (a) Representative roseplots for MDA-MB-231 cells plated on glass, Collagen I, Fibronectin, Tenascin C, or Collagen IV for 16 h and imaged every 10 min. Each line represents an individual cell. Axis length is 500 μ m. Quantification of cell migration speed ( μ m/min) (b) and persistence (c). Results show entire distribution, no ECM (n = 128 cells), Collagen I (n = 45 cells), Fibronectin (n = 91 cells), Tenascin C (n = 25 cells), or Collagen IV (n = 39 cells). Correlation between 2D persistence and 2D cell migration speed (d) and between cell area ( μ m 2 ) and 2D cell migration speed (e). Correlation characterized by r 2 , p value, and Q 2 . (f) Representative roseplots for MDA-MB-468 cells plated on glass, Collagen I, Fibronectin, Tenascin C, or Collagen IV for 16 h and imaged every 10 min. Each line represents an individual cell. The axis length is 500 μ m. Quantification of cell migration speed ( μ m/min) (g) and persistence (h). Results show entire distribution, no ECM (n = 220 cells), Collagen I (n = 68 cells), Fibronectin (n = 126 cells), Tenascin C (n = 24 cells), or Collagen IV (n = 63 cells). Correlation between 2D persistence and 2D cell migration speed (i) and between the cell area and 2D cell migration speed (j). Correlation characterized by r 2 , p value, and Q 2 . Significance determined by one-way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.005, ns is not significant.

Article Snippet: We used the following ECM proteins: Collagen I protein (CB-40236; Fisher Scientific, Hampton, NH), Fibronectin protein (F1141; SIGMA, St. Louis, MO), Tenascin C (R&D systems, 3358TC050), Collagen IV protein (Abcam, ab7536), and Matrigel (growth-factor reduced, Corning, CB-40230C).

Techniques: Migration

Journal: APL Bioengineering

Article Title: Cell shape, and not 2D migration, predicts extracellular matrix-driven 3D cell invasion in breast cancer

doi: 10.1063/1.5143779

Figure Lengend Snippet: ECM-driven 3D invasion does not correlate with effects on 2D cell migration. (a) Representative images of spheroids made from 231-GFP cells embedded in media, Collagen I, Fibronectin, Tenascin C, or Collagen IV gels for 5 days. The scale bar is 200 μ m. (b) Quantification of fold change in 231-GFP spheroid area on day 5 relative to day 1. Data pooled from at least five biological replicates, with three technical triplicate per experiment. *** p < 0.001 by one-way ANOVA and Dunn's multiple comparison test. (c) Correlation between mean fold change in spheroid area and 2D cell migration speed for 231-GFP cells. Correlation characterized by r 2 , p value and Q 2 . (d) Representative images of spheroids made from 468-GFP cells embedded in media, Collagen I, Fibronectin, Tenascin C, or Collagen IV gels for 5 days. Scale bar is 200 μ m. (e) Quantification of fold change in 468-GFP spheroid area on day 5 relative to day 1. Data pooled from at least four biological replicates, with three technical triplicate per experiment. *** p < 0.001 by one-way ANOVA and Dunn's multiple comparison test. (f) Correlation between mean fold change in spheroid area and 2D cell migration speed for 468-GFP. Correlation characterized by r 2 , p value, and Q 2 .

Article Snippet: We used the following ECM proteins: Collagen I protein (CB-40236; Fisher Scientific, Hampton, NH), Fibronectin protein (F1141; SIGMA, St. Louis, MO), Tenascin C (R&D systems, 3358TC050), Collagen IV protein (Abcam, ab7536), and Matrigel (growth-factor reduced, Corning, CB-40230C).

Techniques: Migration, Comparison

Journal: APL Bioengineering

Article Title: Cell shape, and not 2D migration, predicts extracellular matrix-driven 3D cell invasion in breast cancer

doi: 10.1063/1.5143779

Figure Lengend Snippet: Adhesion classifies ECM-driven 2D migration and 3D invasion. (a) Cell adhesion to predict binary classification of 2D cell migration speed of MDA-MB-231 cells on plastic, Collagen I, Fibronectin, Tenascin C, or Collagen IV. (b) AUROC scores of binary AdaBoost classifier models (a) using all 11 cell shape parameters, cell size parameters (area/cell, perimeter, mean radius, min feret diameter, max feret diameter), cell irregularity parameters (solidity, extent, form factor), and cell elongation parameters (eccentricity, aspect ratio, compactness). (c) 2D cell migration to predict binary classification of mean fold change of spheroid area of 231-GFP cells embedded in media, Collagen I, Fibronectin, Tenascin C, or Collagen IV. (d) AUROC scores of binary classifier models (c) using 2D cell migration (cell migration speed and persistence alone or together). (e) Cell adhesion to predict binary classification of mean fold change of spheroid area of 231-GFP cells embedded in media, Collagen I, Fibronectin, Tenascin C, or Collagen IV. (f) AUROC scores of binary classifier models (e) using all 11 cell shape parameters, cell size parameters, cell irregularity parameters, and cell elongation parameters.

Article Snippet: We used the following ECM proteins: Collagen I protein (CB-40236; Fisher Scientific, Hampton, NH), Fibronectin protein (F1141; SIGMA, St. Louis, MO), Tenascin C (R&D systems, 3358TC050), Collagen IV protein (Abcam, ab7536), and Matrigel (growth-factor reduced, Corning, CB-40230C).

Techniques: Migration

Journal: APL Bioengineering

Article Title: Cell shape, and not 2D migration, predicts extracellular matrix-driven 3D cell invasion in breast cancer

doi: 10.1063/1.5143779

Figure Lengend Snippet: ECM-driven predictions are cell-line specific. (a) Representative cell adhesion images show cells fixed and stained with Phalloidin (red) and DAPI (blue) after 2 h on Matrigel. The scale bar is 100 μ m. Quantification of cell migration speed (b) and persistence (c). Data show entire distribution, with no ECM (n = 30), FN (n = 36), TNC (n = 19), Matrigel (n = 36). (d) Quantification of fold change in BT-549-GFP spheroid area on day 5 relative to day 1. Data pooled from 1 biological replicate, with 11 technical triplicate. *** p < 0.001 by one-way ANOVA and Dunn's multiple comparison test. (e) Prediction of 2D cell migration speed, 2D persistence, and 3D invasion of BT-549 cells on Tenascin C and Matrigel from MDA-MB-231, MDA-MB-468, and combined PLS models built with two principal components for MDA-MB-231 model and three principal components for MDA-MB-468 and combined cell lines models. Predictions were done without BT-549 data, with the addition of BT-549 no ECM, and with the addition of BT-549 no ECM and Fibronectin. Numbers represent the actual and predicted values for each metric. Colors represent percent error, where green is a low error and red is a high error value, indicated by the color gradient.

Article Snippet: We used the following ECM proteins: Collagen I protein (CB-40236; Fisher Scientific, Hampton, NH), Fibronectin protein (F1141; SIGMA, St. Louis, MO), Tenascin C (R&D systems, 3358TC050), Collagen IV protein (Abcam, ab7536), and Matrigel (growth-factor reduced, Corning, CB-40230C).

Techniques: Staining, Migration, Comparison