Real-Time Imaging of Notch Activation with a Luciferase Complementation-Based Reporter

Ma. Xenia G. Ilagan,* Sora Lim, Mary Fulbright, David Piwnica-Worms, Raphael Kopan*

*To whom correspondence should be addressed. E-mail: ilaganmg{at}wustl.edu (M.X.G.I.); kopan{at}wustl.edu (R.K.)

This PDF file includes:

• Fig. S1. Transcriptional activation profile of untagged and CLuc-tagged RBPjκ in RBPjκ-null (OT11) fibroblasts.
• Fig. S2. Transcriptional activation profile of untagged and NLuc-tagged Notch receptor variants.
• Fig. S3. Effect of DNA binding and Luc fragment domain swapping on the complementation between NΔE and RBPjκ.
• Fig. S4. LCI reporting of the involvement of MAML in EBNA2-RBPjκ interactions and ternary complex formation with NICD and RBPjκ.
• Fig. S5. Establishment of stable Notch/RBPjκ LCI reporter lines for pathway analyses and high-throughput screening.
• Fig. S6. Determining the rank order of potency of representative g-secretase inhibitors with the NΔE LCI reporter.
• Fig. S7. Monitoring ligand-independent activation of Notch by the Ca²⁺ chelators BAPTA and EDTA using LCI.
• Fig. S8. Monitoring the formation of the NICD-RBPjκ-MAML ternary complex after ligand-dependent activation.
• Fig. S9. Activation of full-length Notch receptors with Dll1-Fc immobilized with antibodies against Fc.
• Fig. S10. Monitoring NotchFL activation by clustered Dll1-Fc with LCI does not depend on downstream transcription and translation.
• Fig. S11. Kinetic and dose-response analyses of S2 and S3 cleavage inhibition in NotchFL LCI reporter cells.
  

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© 2011 American Association for the Advancement of Science