anti py 1068 egfr  (Cell Signaling Technology Inc)

Journal: Theranostics

Article Title: Overcoming resistance to EGFR monotherapy in HNSCC by identification and inhibition of individualized cancer processes

doi: 10.7150/thno.64347

Figure Lengend Snippet: The PaSSS-based drug combinations induced an immune response of PBMC in SCC25 and Cal27 models in vitro and reduced tumor growth in vivo . ( A ) To examine IFN-γ secretion (as illustrated in the scheme on the left) SCC25 cells (middle panel) and Cal27 cells (right panel) were treated with either anti-EGFR monotherapy ( Er ) or the PaSSS-based combination. C stands for control. After 48h and 96h respectively the supernatants were collected for IFN-γ levels quantification (*P < 0.05, **P < 0.001). ( B ) SCC25 (middle panel) and Cal27 cells (right panel) were co-cultured (CC) with PBMCs (as illustrated in the scheme on the left) and treated for 48h and 96h respectively with either anti -EGFR monotherapy or the predicted combination (with or without the addition of 10μg/ml Keytruda, Ky ). PBMCs were then collected and CD3 levels in CD8 positive cells were measured (*P < 0.02 for SCC25,*P < 0.007 for Cal27). ( C ) SCC25 ( C , upper right panel) and Cal27 ( C , lower right panel) were CC with non-activated /activated PBMCs ( AC PBMC ) and then the cells were treated with either anti-EGFR monotherapy or the predicted combination with or without the addition of 10 μg/ml Keytruda for 96h. Cell survival was measured via methylene blue. (D) SCC25 ( D, left panel) or Cal27 ( D , right panel) were injected subcutaneously into mice, and once tumors reached 50 mm , treatments were initiated. In both cases, the PaSSS-based drug combinations (see black arrows) inhibited tumor growth and demonstrated an effect superior to monotherapy of erlotinib or to the drug combinations predicted to partially target the PaSSS (*P < 0.03 for SCC25) (*P < 0.03 for Cal27) (see Figures ,4 for details regarding the altered signaling signatures and the PaSSS-based drug combination predictions). ( E ) Representative, treated and untreated SCC25 and Cal27 tumors, harvested after 25 days and 14 days respectively, are shown. Panels (A-C) were created using BioRender.com.

Article Snippet: The following antibodies were used: anti-pY (1068) EGFR (#3777), anti-PARP (#9542), anti-pS(235/236)S6 (#4858), anti- S6 (#2217), Phospho-4E-BP1 (Thr37/46) (236B4) (#2855), 4E-BP1 (53H11) (#9644), Phospho-Akt (Ser473) (D9E) XP, (#4060), Acetyl-CoA Carboxylase (C83B10)( #3676), Phospho-Acetyl-CoA Carboxylase (Ser79) (#3661), all were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: In Vitro, In Vivo, Control, Cell Culture, Injection

Journal: Theranostics

Article Title: Overcoming resistance to EGFR monotherapy in HNSCC by identification and inhibition of individualized cancer processes

doi: 10.7150/thno.64347

Figure Lengend Snippet: Anti-EGFR monotherapy fails to reduce the unbalanced flux in HNSCC human samples. ( A ) Changes in gene expression levels were acquired from 15 HNSCC patients before and after treatment with cetuximab for 2 weeks as shown in the illustration. ( B ) Patient-specific barcodes were generated for each patient before (left panel) and after the treatment (right panel). Negative/positive amplitude denotes how the patients are correlated with respect to a particular process. For example, patient 1 harbors process 12- (labeled with a black arrow), whereas patient 2 harbors process 12+. Therefore, transcripts that participate in process 12 deviate from the steady state in opposite directions in these patients. ( C ) Only processes in which EGFR participates are shown for each patient. EGFR upregulation or downregulation due to the process were defined as explained in the Figure legend of and labeled with green (upregulation due to a process) or red (downregulation due to a process) colors. In certain tumors, in which a clear reduction in the levels of EGFR and EGFR-related transcripts was detected, other onco-transcripts were upregulated. ( D ) Patient-specific barcodes are shown for patients #1,14,15. EGFR participation in active processes is labeled with green and red colors as indicated. Examples for a change in the experimental gene expression levels in response to EGFR inhibition are shown for selected genes and for each patient in lower panels. (B - barcodes before the treatment ; A - after the treatment ) . Panel (A) was created using BioRender.com.

Article Snippet: The following antibodies were used: anti-pY (1068) EGFR (#3777), anti-PARP (#9542), anti-pS(235/236)S6 (#4858), anti- S6 (#2217), Phospho-4E-BP1 (Thr37/46) (236B4) (#2855), 4E-BP1 (53H11) (#9644), Phospho-Akt (Ser473) (D9E) XP, (#4060), Acetyl-CoA Carboxylase (C83B10)( #3676), Phospho-Acetyl-CoA Carboxylase (Ser79) (#3661), all were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Gene Expression, Generated, Labeling, Inhibition

Journal: Cell Systems

Article Title: Interdependence between EGFR and Phosphatases Spatially Established by Vesicular Dynamics Generates a Growth Factor Sensing and Responding Network

doi: 10.1016/j.cels.2018.06.006

Figure Lengend Snippet: Composite video comprising: EGFR-mCitrine (left), EGF-Alexa647 (middle), and EGFR-mCitrine (magenta)-EGF-Alexa647 (green) overlay. Images were taken at 1-min intervals for ∼120 min after 200 ng/mL EGF-Alexa647 stimulation. Scale bars, 10 μm.

Article Snippet: Rabbit anti EGFR pY 1068 , Cell Signaling Technology , Cat. # 3777.

Techniques:

Journal: Cell Systems

Article Title: Interdependence between EGFR and Phosphatases Spatially Established by Vesicular Dynamics Generates a Growth Factor Sensing and Responding Network

doi: 10.1016/j.cels.2018.06.006

Figure Lengend Snippet: Composite video comprising: EGFR-mCitrine (left), EGF-Alexa647 (middle), and EGFR-mCitrine (magenta)-EGF-Alexa647 (green) overlay. Images were taken at 1-min intervals for ∼120 min after 200 ng/mL EGF-Alexa647 stimulation. Scale bars, 10 μm.

Article Snippet: Rabbit anti EGFR pY 1068 , Cell Signaling Technology , Cat. # 3777.

Techniques:

Journal: Cell Systems

Article Title: Interdependence between EGFR and Phosphatases Spatially Established by Vesicular Dynamics Generates a Growth Factor Sensing and Responding Network

doi: 10.1016/j.cels.2018.06.006

Figure Lengend Snippet:

Article Snippet: Rabbit anti EGFR pY 1068 , Cell Signaling Technology , Cat. # 3777.

Techniques: Modification, Virus, Recombinant, Blocking Assay, cDNA Synthesis, Sequencing, Plasmid Preparation, Expressing, Construct, Software, Magnetic Beads