Journal: Oncogene

Article Title: FGFR3-TACC3 fusion proteins act as naturally occurring drivers of tumor resistance by functionally substituting for EGFR/ERK signaling

doi: 10.1038/onc.2016.216

Figure Lengend Snippet: FGFR3 is activated in FaDu-resistant variant cell lines and maintains ERK signaling upon EGFR blockade. ( a ) Lysates were prepared from FaDu P1, V1 or V2 cells and used to assess tyrosine phosphorylation of 49 human RTKs with the Human Phospho-RTK Array Kit. Active RTKs of note are boxed and labeled. The unlabeled spots on the corners of the membranes are positive controls. ( b ) Lysates from FaDu P1, V1 or V2 cells were subjected to western blot analysis with antibodies against phospho-MET, MET and actin. ( c ) Lysates from FaDu P1, V1 or V2 cells were subjected to immunoprecipitation with anti-phosphotyrosine (p-Tyr) antibody 4G10 conjugated to agarose beads. The presence of FGFR3 and Src (a positive control) in the immunoprecipitates was assessed by western blot analysis. ( d ) FaDu V2 cells were treated for 30 min with control antibody (10 μg/ml) plus vehicle, REGN955 (10 μg/ml), 100 nM PHA665752 (a MET tyrosine kinase inhibitor) or the combination of REGN955 plus PHA665752. Following treatment, cell lysates were subjected to western blot analysis with antibodies against phospho-ERK, ERK, phospho-MET and MET. ( e ) FaDu V2 cells were treated for 1 h with vehicle or with 25 nM AZD4547, a pan-FGFR tyrosine kinase inhibitor. Following treatment, cell lysates were subjected to immunoprecipitation with p-Tyr antibody 4G10 conjugated to agarose beads. The presence of FGFR3 and Src in the immunoprecipitates was assessed by western blot analysis. ( f ) FaDu V1 or V2 cells were treated for 30 min with control antibody (10 μg/ml) plus vehicle, REGN955 (10 μg/ml), 25 nM AZD4547 or the combination of REGN955 plus AZD4547. Following treatment, cell lysates were subjected to western blot analysis with antibodies against phospho-ERK, ERK, phospho-AKT and AKT.

Article Snippet: Cells were treated with the following reagents (doses and treatment times are indicated in the Figure Legends): REGN1400 (ERBB3-blocking antibody), REGN955 (EGFR-blocking antibody), MEK inhibitor GSK1120212 (Selleckchem, Houston, TX, USA), FGFR TKI AZD4547 (Selleckchem), MET TKI PHA665752 (Sigma, St Louis, MO, USA), EGFR TKI AZD9291 (Selleckchem), PI3K inhibitor BYL719 (Selleckchem), human NRG1 (R&D Systems, Minneapolis, MN, USA) and human EGF (R&D Systems).

Techniques: Variant Assay, Phospho-proteomics, Labeling, Western Blot, Immunoprecipitation, Positive Control, Control

Journal: Genes & Cancer

Article Title: Correlation between c-Met and ALDH1 contributes to the survival and tumor-sphere formation of ALDH1 positive breast cancer stem cells and predicts poor clinical outcome in breast cancer

doi: 10.18632/genesandcancer.148

Figure Lengend Snippet: List of c-Met inhibitors

Article Snippet: c-Met inhibitors (AMG-208, BMS 777607, Cabozantinib, Crizotinib, Foretinib, NVP-BVU972, PF-04217903, PHA-665752, Tivantinib) were purchased from Namiki Inc. (Japan).

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: Autism Spectrum Disorder Risk Factor Met Regulates the Organization of Inhibitory Synapses

doi: 10.3389/fnmol.2021.659856

Figure Lengend Snippet: Hepatocyte growth factor (HGF) increases gephyrin clustering in vitro . Hippocampal neurons were left untreated or were treated with DMSO, HGF, or PHA-665752. Representative images of gephyrin- or MAP2-stained hippocampal neurons are shown in the absence (A–A”) or presence (B–B”) of HGF. Overview images are shown in ( A,B ; scale bar 20 μm) while insets are enlarged in ( A’,A”,B’,B” ; scale bar 10 μm). Panels (A’,B’) depict confocal images, (A”,B”) processed images for the quantification of gephyrin cluster (red) densities within masks of MAP2-positive dendritic segments (25 μm). ( C) Quantification of gephyrin cluster densities. One-way ANOVA and Tukey’s multiple comparison test F (4,364) = 0.7245. CTR: n = 75, HGF: n = 71, DMSO n = 78, PHA n = 73, PHA/HGF n = 72. *** p < 0.0001, error bars SEM. (D,E) Untreated hippocampal neurons (D) or treated with HGF (E) were stained for phosphorylated p70S6K (pS6K, scale bar 20 μm). Insets are enlarged in ( D’,E’ ; scale bar 10 μm) for determination of mean fluorescence intensities of phosphorylated p70S6K signals within a mask of MAP2-positive cell bodies. Dashed outlines mark the area of the cell bodies. (F) Quantification of phosphorylated p70S6K intensities. CTR n = 57, DMSO n = 83, HGF n = 83, PHA n = 68, PHA/HGF n = 77. One-way ANOVA and Tukey’s multiple comparison test F (4,363) = 3.369. *** p < 0.0001, error bars: SEM.

Article Snippet: Inhibitors were dissolved in dimethylsulfoxide (DMSO), HGF (Merck KGaA, Darmstadt, Germany) in PBS and added to hippocampal neurons (DIV 9) to final concentrations of 50 ng/ml for HGF, 200 nm for mTOR inhibitor rapamycin (Merck), and 1 μM PHA-665752 (Merck KGaA, Darmstadt, Germany).

Techniques: In Vitro, Staining, Fluorescence