tyrosine 416 residue  (Cell Signaling Technology Inc)

Journal: BMC Cancer

Article Title: Insulin growth factor 1 like receptor (IGF-1R)

doi: 10.1186/s12885-016-2796-x

Figure Lengend Snippet: IGF-1R increase in resistant cells upon exposure to anti-EGFR antibody cetuximab. a UMSCC6 cetuximab resistant and sensitive cell lines at passage 17 were exposed to increasing concentration of cetuximab to measure cell viability by absorbance in a crystal violet assay (■) Resistant cells in the absence of cetuximab, (●) sensitive cells in the presence of cetuximab (▲) resistant cells in the presence of cetuximab. b Representative blot of total EGFR and IGF-1R show increased levels in cetuximab resistant cells relative to parental cells. c , d Quantitation of total EGFR and IGF-1R levels normalized to GAPDH from four independent experiments

Article Snippet: Antibodies against total EGFR, IGF1R, Src, phospho-EGFR tyrosine 845, phospho-Src tyrosine 416, GAPDH and mouse immunoprecipitation (IP) specific antibody were obtained from Cell Signaling Technology (Beverly, MA).

Techniques: Concentration Assay, Crystal Violet Assay, Quantitation Assay

Journal: BMC Cancer

Article Title: Insulin growth factor 1 like receptor (IGF-1R)

doi: 10.1186/s12885-016-2796-x

Figure Lengend Snippet: Phosphorylation of EGFR Y845 is not inhibited with Src inhibitors PP1 and Dasatinib. a EGFR Y845 levels show increased levels in response to cetuximab with time in resistant cells ( left panel ) and constant levels in sensitive cells ( right panel ). b Quantitation of EGFR Y845 normalized to GAPDH from four independent experiments after increasing times of exposure to10 μM of PP1. c phospho-Src Y416 show increased levels in response to cetuximab with time in resistant cells ( left panel ) and constant levels in sensitive cells ( right panel ). d Quantitation of p-Src Y416 normalized to GAPDH from four independent experiments after increasing times of exposure to cetuximab. e Representative western blot of EGFR Y845 show constant levels in response to PP1 in both resistant cells ( left panel ) and sensitive cells ( right panel ) in presence of cetuximab. f Representative western blot of p-Src 416 show reducing levels in response to PP1 in both resistant cells ( left panel ) and sensitive cells ( right panel ) with increasing time. g As in E above but after exposure to Dasatinib. h As in E above but after exposure to Dasatinib. i Proliferation of resistant and sensitive cell lines after exposure to PP1 for 72 h, measured using the CCK-8 assay

Article Snippet: Antibodies against total EGFR, IGF1R, Src, phospho-EGFR tyrosine 845, phospho-Src tyrosine 416, GAPDH and mouse immunoprecipitation (IP) specific antibody were obtained from Cell Signaling Technology (Beverly, MA).

Techniques: Phospho-proteomics, Quantitation Assay, Western Blot, CCK-8 Assay

Journal: BMC Cancer

Article Title: Insulin growth factor 1 like receptor (IGF-1R)

doi: 10.1186/s12885-016-2796-x

Figure Lengend Snippet: Knockdown of IGF-1R reduces the proliferation of resistant cells. a Relative expression of EGFR and IGF1R in resistant cells subjected to knockdown with three independent siRNAs targeting EGFR or IGF1R respectively after 60 h. Scrambled control was set to 1. b as in A but for the sensitive cell line. c Representative western blot indicating reduced levels of EGFR (left panel) or IGF1R (right panel) after knockdown with siRNA #1 and #8 respectively in resistant cells. GAPDH was used as a loading control protein was quantified at 60 h. d As in C but for the sensitive cell line. e Proliferation of resistant and sensitive cell lines in untreated, scrambled and specific knockdown conditions, measured using the CCK-8 assay, measured at 72 h. IGF1R has a greater effect on proliferation in resistant as compared to sensitive cells

Article Snippet: Antibodies against total EGFR, IGF1R, Src, phospho-EGFR tyrosine 845, phospho-Src tyrosine 416, GAPDH and mouse immunoprecipitation (IP) specific antibody were obtained from Cell Signaling Technology (Beverly, MA).

Techniques: Knockdown, Expressing, Control, Western Blot, CCK-8 Assay