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: Cancer Management and Research

Article Title: Progress toward overcoming hypoxia-induced resistance to solid tumor therapy

doi: 10.2147/CMAR.S58285

Figure Lengend Snippet: Examples of drugs targeting multiple pathways associated with hypoxia and hypoxia inducible factor 1 (HIF-1)

Article Snippet: Signaling , MET kinase , PHA665752 , Funakoshi et al, Puri et al .

Techniques: Activity Assay, Binding Assay