Journal: International Journal of Molecular Sciences

Article Title: EGF Potentiation of VEGF Production Is Cell Density Dependent in H292 EGFR Wild Type NSCLC Cell Line

doi: 10.3390/ijms151017686

Figure Lengend Snippet: VEGF-A production in confluent vs . subconfluent cells treated with EGF. ( A ) Confluent cells exhibit dose dependent increase in VEGF-A production in response to EGF whereas subconfluent cell VEGF-A production was insensitive to EGF treatment; and ( B ) Basal VEGF-A production in confluent cells is greater than that in subconfluent cells and is significantly stimulated by EGF while 30 min pre-incubation with Cetuximab prevented EGF stimulated VEGF-A production. * p < 0.05 Student’s t -test, n.s. = not significant, n = 9.

Article Snippet: EGFR, cMet, and VEGF expression/phosphorylation kits were provided by Meso Scale Discovery (Rockville, MD, USA).

Techniques: Incubation

Journal: International Journal of Molecular Sciences

Article Title: EGF Potentiation of VEGF Production Is Cell Density Dependent in H292 EGFR Wild Type NSCLC Cell Line

doi: 10.3390/ijms151017686

Figure Lengend Snippet: STAT3, but not PI3K, contributes to EGFR-mediated VEGF-A production in confluent cells. ( A ) R&D Systems phospho-protein dot-blot of membranes incubated with equal amounts of cell lysate prepared 30 min after 100 ng/mL EGF treatment on cells seeded as confluent or subconfluent. Arrow indicates STAT3-ser727 phosphorylation; ( B ) VEGF-A production in confluent cells 24 h after indicated treatment, Ctxmab = Cetuximab, * p < 0.05, one-way ANOVA with Bonferonni post test; and ( C ) VEGF-A production in confluent cells 24 h after indicated treatment. * p < 0.05, students t -test vs . SP1.

Article Snippet: EGFR, cMet, and VEGF expression/phosphorylation kits were provided by Meso Scale Discovery (Rockville, MD, USA).

Techniques: Dot Blot, Incubation, Phospho-proteomics

Journal: Molecular Vision

Article Title: Glucosamine inhibits epidermal growth factor-induced proliferation and cell-cycle progression in retinal pigment epithelial cells

doi:

Figure Lengend Snippet: Effects of glucosamine (GlcN) on transactivation of epidermal growth factor receptor (EGFR). Human retinal pigment epithelial cell line (ARPE-19) cells were treated with 2.5 mM or 5.0 mM GlcN, or 30 mM glucose for 24 h, stimulated with 10 ng/ml EGF, and stained with PE-conjugated anti-total-EGFR antibody and Alexa-Fluor-647-conjugated anti-p-EGFR antibody (Y845). The levels of phosphorylated ( A ) and total EGFR ( B ) were analyzed by flow cytometry. The data are representative of at least three independent experiments. Cells cultured under serum-free conditions were used as the control. C : western blot analysis of ARPE-19 cells cultured to 80% confluence; treated with 2 μg/ml tunicamycin, 30 mM glucose, or 5 mM GlcN in serum-free medium for a further 24 h; then stimulated with EGF for 5 min.

Article Snippet: The following primary antibodies were used: phycoerythrin (PE)-conjugated anti-total-EGFR (cells were not permeabilized for total EGFR detection) and Alexa-Fluor-647-conjugated anti-p-EGFR (Y845; BD PharMingen).

Techniques: Staining, Flow Cytometry, Cell Culture, Western Blot

Journal: Cell

Article Title: EGFR ligands differentially stabilize receptor dimers to specify signaling kinetics

doi: 10.1016/j.cell.2017.09.017

Figure Lengend Snippet: (A) Representative time-courses of EGFR phosphorylation at Y1173, Y845, and Y1086 in MCF-7 cells induced by saturating levels of EGF (16 nM), EREG (20 μM) or EPGN (20 μM). Anti-Grb2 is used as loading control. Data for pY1173 were generated by stripping and reprobing pY845 blots, so use the same loading controls.

Article Snippet: Total Grb2 levels are included as a loading control. ( B ) Upper panel: Western blot comparing time-courses in MCF-7 cells over 1 h for EGFR Y1086 phosphorylation induced by epigen (at 1 mM and 20 μM) and EGF (16 nM).

Techniques: Phospho-proteomics, Control, Generated, Stripping Membranes