egf (final concentration, 30 ng/ml or 300 ng/ml as listed in various figure legends)  (R&D Systems)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Motility-limited aggregation of mammary epithelial cells into fractal-like clusters

doi: 10.1073/pnas.1905958116

Figure Lengend Snippet: Jamming-like phase diagram based on neighbor density and EGF. (A) Neighbor density is a readout of individual, clustered, and spanning phases. Each line indicates data from a representative well treated with 0.075 ng/mL EGF, for various initial starting densities with several replicates shown. (Right) Reference snapshots of live cells (GFP cytoplasm, gray) for each of the density regimes. (B) A jamming-like phase diagram defined with neighbor density and EGF concentration. Gray circles indicate motile cells (>7 μm/h) and Xs indicate nonmotile cells (<7 μm/h) at experimentally tested EGF concentrations, while cell speed is interpolated across untested values. Replicate conditions for each EGF concentration and initial starting density were used to construct the phase diagram. (C) Clustering and spanning are captured by a computational model of self-propelled particles with varying polarization force (cell speed) at fixed adhesion (A = 0.03). (Right) Red particles represent individuals, while gray particles are associated with a cluster. (D) A jamming-like phase diagram defined with neighbor density and cell polarization parameter recapitulates the behaviors observed in the experimental low-EGF regime, highlighted in purple in B. Each pixel represents the average values (speed, neighbor number) for a distinct simulation.

Article Snippet: For these experiments, cells were cultured on collagen-coated 96-well plates at varying EGF concentrations (0 to 20 ng/mL) with DMSO and OHT, with a total media volume of 125 μ L. No changes in media color were observed after 60 h. Cells tested negative for Mycoplasma contamination (PlasmoTest; Invivogen) and an absence of additional cell culture contaminants was routinely verified by culturing cells in antibiotic-free media.

Techniques: Concentration Assay, Construct

Journal: International Journal of Bioprinting

Article Title: Tissue-engineered edible bird’s nests (TeeBN)

doi: 10.18063/ijb.691

Figure Lengend Snippet: Preparation and evaluation of the receiving layer of TeeBN. (A) Schematic of receiving layer. Inset images showed the positional relationship: The blue upper layer is the feeding layer, and the white lower layer indicated by the orange arrow is the receiving layer. The table showed the main components of the receiving layer. (B) Dot blotting analysis of the binding between HA with different molecular weight and EGF with different concentration. (C) The EFG level after enzyme digested. **** P < 0.0001, n.s. means not significant (n = 5). (D) SEM image of EBN and (E) TeeBN. Scale bar, 100 μm. (F) The total protein content. ** P < 0.01 and **** P < 0.0001 (n = 5). (G) The total EGF content. **** P < 0.0001 (n = 5). (H) The total sialic acid content. **** P < 0.0001 (n = 5). (I) The total nitrite content. **** P < 0.0001 (n = 5). (J) The heavy metal As (J), Pb (K), and Cu (L) content. ** P < 0.01 and **** P < 0.0001 (n = 3). All results were analyzed with one-way ANOVA and Tukey’s post-hoc multiple comparisons tests in GraphPad Prism 8 and expressed as mean ± SD. Results of TeeBN vs. EBN-V and TeeBN vs. EBN-I are show in Figures F to L.

Article Snippet: Mouse EGF concentrations were determined by the enzyme-linked immunosorbent assay kit (ELISA; Cusabio, Wuhan, China).

Techniques: Binding Assay, Molecular Weight, Concentration Assay

Journal: Tomography

Article Title: Short-Term Environmental Conditioning Enhances Tumorigenic Potential of Triple-Negative Breast Cancer Cells

doi: 10.18383/j.tom.2019.00019

Figure Lengend Snippet: Epidermal growth factor (EGF) conditioning enhances onset of breast tumor formation. We conditioned MDA-MB-231 cells with EGF (30 ng/mL), FBS, or control for 4 hours before orthotopic implantation of 10 3 cells per mammary fat pad (n = 6 mice per condition). Graph displays tumor incidence over time for each group (A). * P = .048 for EGF versus control and *** P = .0006 for FBS versus control tumor onset by 1-way ANOVA. Mean ± SEM for photon flux over time in each group (B). Note log scale for total flux values. Scatterplot shows mean ± SEM for final tumor volume on day 46 measured in 3 dimensions with calipers (C). ** P = .004 for FBS versus control tumor volume by Kruskal–Wallis test.

Article Snippet: The next day, we conditioned breast cancer cells by adding FBS (final concentration, 10%), EGF (final concentration, 30 ng/mL or 300 ng/mL as listed in various figure legends) (R&D Systems, Minneapolis, MN), ridaforolimus (Selleck Chemicals, Houston, TX; final concentration, 10 nM), or trametinib (Selleck Chemicals; final concentration, 100 nM) to their existing media and incubated for 4 hours.

Techniques:

Journal: Tomography

Article Title: Short-Term Environmental Conditioning Enhances Tumorigenic Potential of Triple-Negative Breast Cancer Cells

doi: 10.18383/j.tom.2019.00019

Figure Lengend Snippet: Tumor Formation after Bilateral Injection of MDA-MB-231 Cells after Conditioning with EGF Compared to Control or FBS Conditioned Cells a

Article Snippet: The next day, we conditioned breast cancer cells by adding FBS (final concentration, 10%), EGF (final concentration, 30 ng/mL or 300 ng/mL as listed in various figure legends) (R&D Systems, Minneapolis, MN), ridaforolimus (Selleck Chemicals, Houston, TX; final concentration, 10 nM), or trametinib (Selleck Chemicals; final concentration, 100 nM) to their existing media and incubated for 4 hours.

Techniques: Injection

Journal: Tomography

Article Title: Short-Term Environmental Conditioning Enhances Tumorigenic Potential of Triple-Negative Breast Cancer Cells

doi: 10.18383/j.tom.2019.00019

Figure Lengend Snippet: Conditioning treatments do not alter population-level proliferation or adhesion of cancer cells in cell-based assays. We conditioned MDA-MB-231 cells with EGF (30 ng/mL), ridaforolimus (100 nM), trametinib (100 nM), FBS, or control prior to seeding 10 3 cells per well in a 96-well plate (n ≥ 4 per condition) (A). We normalized bioluminescence on days one and two to corresponding values on day 0 for each group. Graph shows mean + SEM for normalized bioluminescence on day 2 for each condition as a marker of proliferation. * P = .0261 for control versus trametinib by 1-way ANOVA. We conditioned cells with the same treatments listed in (A) and then seeded 2.5 × 10 5 cells per well onto confluent monolayers of human mammary fibroblasts (HMFs) in a 24-well plate (B). We washed off nonadherent cells with PBS after 15 minutes and then quantified the number of adherent breast cancer cells. Graph shows mean + SEM for cells adhering to breast cancer cells for each condition (EGF, 30 ng/mL; ridaforolimus, 100 nM; trametinib, 100 nM; FBS; or control) (n ≥ 10 per condition).

Article Snippet: The next day, we conditioned breast cancer cells by adding FBS (final concentration, 10%), EGF (final concentration, 30 ng/mL or 300 ng/mL as listed in various figure legends) (R&D Systems, Minneapolis, MN), ridaforolimus (Selleck Chemicals, Houston, TX; final concentration, 10 nM), or trametinib (Selleck Chemicals; final concentration, 100 nM) to their existing media and incubated for 4 hours.

Techniques: Marker

Journal: Tomography

Article Title: Short-Term Environmental Conditioning Enhances Tumorigenic Potential of Triple-Negative Breast Cancer Cells

doi: 10.18383/j.tom.2019.00019

Figure Lengend Snippet: FBS conditioning promotes tumor formation by patient-derived Vari-068 breast cancer cells. We conditioned Vari-068 breast cancer cells with EGF (300 ng/mL), FBS, or control for 4 hours before implanting 10 3 (left) or 10 4 (right) cells per mammary fat pad (n = 3 mice per condition). Graphs show incidences of tumor formation for each condition and amount of injected cells (A). Bioluminescence imaging for each condition over time expressed as mean ± SEM (B). Scatterplots with mean ± SEM for final tumor volumes measured by calipers on day 78 (C). * P = .017 for FBS versus control by Kruskal–Wallis test.

Article Snippet: The next day, we conditioned breast cancer cells by adding FBS (final concentration, 10%), EGF (final concentration, 30 ng/mL or 300 ng/mL as listed in various figure legends) (R&D Systems, Minneapolis, MN), ridaforolimus (Selleck Chemicals, Houston, TX; final concentration, 10 nM), or trametinib (Selleck Chemicals; final concentration, 100 nM) to their existing media and incubated for 4 hours.

Techniques: Derivative Assay, Injection, Imaging