Supplementary Materials

Supplementary Materials for:

Systematic identification of signaling pathways with potential to confer anticancer drug resistance

Colin A. Martz, Kathleen A. Ottina, Katherine R. Singleton, Jeff S. Jasper, Suzanne E. Wardell, Ashley Peraza-Penton, Grace R. Anderson, Peter S. Winter, Tim Wang, Holly M. Alley, Lawrence N. Kwong, Zachary A. Cooper, Michael Tetzlaff, Pei-Ling Chen, Jeffrey C. Rathmell, Keith T. Flaherty, Jennifer A. Wargo, Donald P. McDonnell, David M. Sabatini,* Kris C. Wood*

*Corresponding author. E-mail: sabatini{at}wi.mit.edu (D.M.S.); kris.wood{at}duke.edu (K.C.W.)

This PDF file includes:

  • Fig. S1. Schematic of the screen.
  • Fig. S2. Stable transfection of constructs.
  • Fig. S3. Results of a screen in UACC-62 melanoma cells (BRAFV600E) for pathways conferring resistance to MEK1/2 inhibitor AZD6244.
  • Fig. S4. Meta-analysis of screening results across 13 targeted therapies.
  • Fig. S5. Resistance to etoposide mediated by stable expression of non-cleavable caspases in MCF-7 breast cancer cells.
  • Fig. S6. Immunoblotting of BRAFV600E melanoma cells expressing pathway-activating constructs and treated with MAPK pathway inhibitors.
  • Fig. S7. Resistance to targeted and cytotoxic drugs in breast cancer cells treated with a soluble Notch agonist.
  • Fig. S8. Differentiation markers and signaling in MCF-7 breast cancer cells expressing Notch1.
  • Fig. S9. Characterization of dedifferentiation-associated phenotypes in breast cancer cells with activated Notch1.
  • Fig. S10. Analysis of Notch1 pathway members and EMT markers in a mouse model of tamoxifen-resistant breast cancer (TamR).
  • Fig. S11. Analysis of Notch1 pathway members and EMT markers in human breast cancer patients.
  • Fig. S12. Estrogen receptor–driven resistance to MAPK inhibitors.
  • Fig. S13. Notch1 target gene expression in melanoma cells with activated Notch1.
  • Fig. S14. Western blot analysis of differentiation markers and signaling in BRAF-mutant melanoma cells expressing Notch1.
  • Fig. S15. Characterization of dedifferentiation-associated phenotypes in melanoma cells with activated Notch1.
  • Fig. S16. Notch1 hairpin validation.
  • Fig. S17. Characterization of evolved MAPK inhibitor–resistant, BRAF-mutant melanoma cell lines.
  • Fig. S18. Analysis of patient tumors in cohort 1.
  • Fig. S19. Analysis of patient tumors in cohort 2.
  • Fig. S20. Resistance pathway inhibitors sensitize intrinsically resistant melanoma cells to VX-11E independently of inhibitor effects on cell viability.
  • Fig. S21. Vector maps for vectors used in this study.
  • Legends for tables S1 to S7
  • Legend for date file S1

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Other Supplementary Material for this manuscript includes the following:

  • Table S1 (Microsoft Excel format). List of pathway-activating constructs and controls used in this study.
  • Table S2 (Microsoft Excel format). List of all drugs, drug concentrations, and cell lines screened.
  • Table S3 (Microsoft Excel format). Results of primary screens.
  • Table S4 (Microsoft Excel format). Characterization of cell lines and clonal derivates with evolved resistance to MAPK inhibitors.
  • Table S5 (Microsoft Excel format). Metastatic melanoma patient characteristics (cohorts 1 and 2).
  • Table S6 (Microsoft Excel format). List of attB1/B2 primers used to barcode and amplify constructs by PCR.
  • Table S7 (Microsoft Excel format). Sequences of additional primers used in this study.
  • Data file S1 (Microsoft Word format). Nucleotide sequences.

[Download Tables S1 to S7]

[Download Data File S1]


Citation: C. A. Martz, K. A. Ottina, K. R. Singleton, J. S. Jasper, S. E.Wardell, A. Peraza-Penton, G. R. Anderson, P. S. Winter, T. Wang, H. M. Alley, L. N. Kwong, Z. A. Cooper, M. Tetzlaff, P.-L. Chen, J. C. Rathmell, K. T. Flaherty, J. A. Wargo, D. P. McDonnell, D. M. Sabatini, K. C. Wood, Systematic identification of signaling pathways with potential to confer anticancer drug resistance. Sci. Signal. 7, ra121 (2014).

© 2014 American Association for the Advancement of Science