Receptor Signaling: RTK-Mediated Signaling

ACEA Biosciences xCELLigence

Endogenous Receptor Tyrosine Kinase Short Term Response

Application_Book_06_01
Cells expressing recombinant PDGFRb were seeded on an E-plate 96 at 20,000 cells per well and after overnight growth the medium was replaced with serum free medium (not shown). (A) After 2 hours cells were treated with increasing doses of the PDGF nhibitor Imatinib, incubated for one hour, then stimulated with 10 ng/ml PDGF BB. (B) Imatinib showed a dose-responsive inhibition of the morphology change resulting from PDGF stimulation, with an IC50 value of 135 nM as determined using the xCELLigence software (Data and figures adapted from ACEA Biosciences, unpublished data).

Endogenous Receptor Tyrosine Kinase Expressing Cell Line Response to Inhibitor

Application_Book_06_02(A) Cells expressing constitutively active cMET were seeded on an E-plate 384 at 10,000 cells per well and after overnight growth cells were treated with increasing doses of the cMET inhibitor Crizotinib. (B) Crizotinib showed a doseresponsive morphology change resulting from blocking the constitutive cMET signal, with an IC50 value of 87.5 nM as determined using the xCELLigence software (Data and figures adapted from ACEA Biosciences, unpublished data).

Key Benefits 
  1. The integrated, global response of cells to RTK modulation can be monitored.
  2. Rapid responses involving cell morphology and attachment changes may be assayed on a minute timescale.
  3. Slower responses involving cell proliferation and viability may be assayed over days or weeks.
  4. Receptor activity can often be assessed in the endogenous context, in a disease-relevant cell type.
RTK-Mediated Cell Signaling Supporting Information:

  • RTK-Mediated Cell Signaling Publications
  1. Benzothiophene containing Rho kinase inhibitors: Efficacy in an animal model of glaucoma. Davis RL, Kahraman M, Prins TJ, Beaver Y, Cook TG, Cramp J, Cayanan CS, Gardiner EM,McLaughlin MA, Clark AF, Hellberg MR, Shiau AK, Noble SA, Borchardt AJ. Bioorganic & Medicinal Chemistry Letters. 2010 Jun 1;20(11):3361-6.
  2. Modeling ERBB receptor-regulated G1/S transition to find novel targets for de novo trastuzumab resistance. Sahin O, Fröhlich H, Löbke C, Korf U, Burmester S, Majety M, Mattern J, Schupp I,Chaouiya C, Thieffry D, Poustka A, Wiemann S, Beissbarth T, Arlt D. BMC Syst Biol. 2009 Jan 1;3:1.
  3. Label-free and real-time cell-based kinase assay for screening selective and potent receptor tyrosine kinase inhibitors using microelectronic sensor array. Atienza JM, Yu N, Wang X, Xu X, Abassi Y. J Biomol Screen. 2006 Sep;11(6):634-43.