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Sci. Signal., 10 January 2012
[DOI: 10.1126/scisignal.2002274]

Supplementary Materials for:

Wnt/β-Catenin Signaling and AXIN1 Regulate Apoptosis Triggered by Inhibition of the Mutant Kinase BRAFV600E in Human Melanoma

Travis L. Biechele, Rima M. Kulikauskas, Rachel A. Toroni, Olivia M. Lucero, Reyna D. Swift, Richard G. James, Nick C. Robin, David W. Dawson, Randall T. Moon,* Andy J. Chien*

*To whom correspondence should be addressed. E-mail: andchien{at}uw.edu (A.J.C.); rtmoon{at}uw.edu (R.T.M.)

This PDF file includes:

  • Fig. S1. BRAF and other members of the MAPK family are identified as regulators of Wnt/β-catenin signaling in melanoma cells.
  • Fig. S2. BRAF is a negative regulator of Wnt/β-catenin signaling and inhibition of BRAF decreases the phosphorylation of β-catenin at Ser33/Ser37/Thr41 in melanoma cells.
  • Fig. S3. Isobologram analysis of WNT3A and PLX4720 shows a synergistic activation of Wnt/β-catenin signaling in melanoma cells.
  • Fig. S4. BRAF inhibition reduces the abundance of phosphorylated Ser33/Ser37/Thr41 β-catenin with no effect on total β-catenin abundance.
  • Fig. S5. Activation of Wnt/β-catenin signaling cooperates with BRAF inhibition to inhibit proliferation of melanoma cells in vivo.
  • Fig. S6. Isobologram analysis of WNT3A and PLX4720 shows a synergistic inhibition of melanoma cell viability.
  • Fig. S7. Activation of Wnt/β-catenin signaling combined with siRNA-mediated knockdown of BRAF promotes apoptosis of melanoma cells.
  • Fig. S8. BRAF inhibition does not regulate AXIN1 mRNA abundance.
  • Fig. S9. siRNA targeting AXIN1 effectively reduces AXIN1 protein abundance.
  • Table S1. siRNA sequences used for the described studies.
  • Table S2. Allelic status of BRAFV600E in different melanoma cell lines.
  • Database S1. A375 melanoma cells expressing the BAR reporter were screened using siRNAs for kinases that regulate Wnt/β-catenin signaling.
  • Database S2. Screen data from database S1 without resazurine normalization.

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Other Supplementary Material for this manuscript includes the following:

  • Database S1 (Microsoft Excel format). A375 melanoma cells expressing the BAR reporter were screened using siRNAs for kinases that regulate Wnt/β-catenin signaling.
  • Database S2 (Microsoft Excel format). Screen data from database S1 without resazurine normalization.
[Download Databases S1 to S2 (Compressed)]

Technical Details

Format: Microsoft Excel

Size: 92 KB (compressed); 95 KB (decompressed)


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Citation: T. L. Biechele, R. M. Kulikauskas, R. A. Toroni, O. M. Lucero, R. D. Swift, R. G. James, N. C. Robin, D. W. Dawson, R. T. Moon, A. J. Chien, Wnt/β-Catenin Signaling and AXIN1 Regulate Apoptosis Triggered by Inhibition of the Mutant Kinase BRAFV600E in Human Melanoma. Sci. Signal. 5, ra3 (2012).

© 2012 American Association for the Advancement of Science


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