Supplementary Materials

Supplementary Materials for:

ATF4 induction through an atypical integrated stress response to ONC201 triggers p53-independent apoptosis in hematological malignancies

Jo Ishizawa, Kensuke Kojima, Dhruv Chachad, Peter Ruvolo, Vivian Ruvolo, Rodrigo O. Jacamo, Gautam Borthakur, Hong Mu, Zhihong Zeng, Yoko Tabe, Joshua E. Allen, Zhiqiang Wang, Wencai Ma, Hans C. Lee, Robert Orlowski, Dos D. Sarbassov, Philip L. Lorenzi, Xuelin Huang, Sattva S. Neelapu, Timothy McDonnell, Roberto N. Miranda, Michael Wang, Hagop Kantarjian, Marina Konopleva, R. Eric. Davis,* Michael Andreeff*

*Corresponding author. E-mail: mandreef{at}mdanderson.org (M.A.) redavis1{at}mdanderson.org (R.E.D.)

This PDF file includes:

  • Fig. S1. Cell cycle changes and DNA damage induced by ONC201.
  • Fig. S2. Effect of ONC201 on primary MCL and AML cells and normal bone marrow cells.
  • Fig. S3. Efficacy of ONC201 in bortezomib-resistant MM cells and combination therapy with ibrutinib in primary MCL cells.
  • Fig. S4. Apoptosis induced by ONC201 in caspase-8– or Bax/Bak-deficient cells.
  • Fig. S5. Changes in the AKT/ERK/FOXO3a pathway with ONC201 treatment.
  • Fig. S6. Immunoblot and PCR analysis of ER stress–related molecules.
  • Fig. S7. Quantification of the immunoblots in Fig. 5 (A and B).
  • Fig. S8. Effects of ER stress inducers on GCN2, eIF2α, ATF4, and mTORC1.
  • Fig. S9. Immunoblot analysis of ISR-related molecules after amino acid or glucose deprivation in OCI-AML3 and JeKo-1 cells.
  • Fig. S10. ATF4 induction by ONC201 or tunicamycin in OCI-AML3 cells with eIF2α knockdown.
  • Fig. S11. ATF4 expression over time in response to ONC201 in OCI-AML3 and JeKo-1 cells.
  • Fig. S12. Effects of CHOP knockdown on ONC201-induced apoptosis.
  • Fig. S13. Effects of ONC201 on IRE1α.
  • Fig. S14. The role of ATF4 in mTORC1 inhibition by ONC201.
  • Fig. S15. Quantification of the immunoblots in Fig. 7A.
  • Table S1. Clinical information for the primary AML samples.
  • Table S2. Summary of changes in the GCN2/eIF2α/ATF4 axis and mTORC1 targets by tunicamycin, amino acid or glucose deprivation, and ONC201 in AML and MCL cells.
  • Table S3. Combination index of each cell line treated with ONC201 and ABT-199.
  • Table S4. shRNA sequences used.
  • Table S5. Antibodies for immunoblot analysis.
  • Table S6. Primers used for PCR.
  • Legends for data files S1 to S3

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Other Supplementary Material for this manuscript includes the following:

  • Data file S1 (Microsoft Excel format). Genes with most positive slope and r in Fig. 4B.
  • Data file S2 (Microsoft Excel format). Genes with most negative slope and r in Fig. 4B.
  • Data file S3 (Microsoft Excel format). Common up-regulated genes in ONC201-sensitive MCL cell lines.

[Download Data Files S1 to S3]


Citation: J. Ishizawa, K. Kojima, D. Chachad, P. Ruvolo, V. Ruvolo, R. O. Jacamo, G. Borthakur, H. Mu, Z. Zeng, Y. Tabe, J. E. Allen, Z. Wang, W. Ma, H. C. Lee, R. Orlowski, D. D. Sarbassov, P. L. Lorenzi, X. Huang, S. S. Neelapu, T. McDonnell, R. N. Miranda, M. Wang, H. Kantarjian, M. Konopleva, R. E. Davis, M. Andreeff, ATF4 induction through an atypical integrated stress response to ONC201 triggers p53- independent apoptosis in hematological malignancies. Sci. Signal. 9, ra17 (2016).

© 2016 American Association for the Advancement of Science