Supplementary Materials

Supplementary Material for:

HIF activation causes synthetic lethality between the VHL tumor suppressor and the EZH1 histone methyltransferase

Abhishek A. Chakraborty, Eijiro Nakamura, Jun Qi, Amanda Creech, Jacob D. Jaffe, Joshiawa Paulk, Jesse S. Novak, Kshithija Nagulapalli, Samuel K. McBrayer, Glenn S. Cowley, Javier Pineda, Jiaxi Song, Yaoyu E. Wang, Steven A. Carr, David E. Root, Sabina Signoretti, James E. Bradner, William G. Kaelin Jr.*

*Corresponding author. Email: william_kaelin{at}dfci.harvard.edu

Published 12 July 2017, Sci. Transl. Med. 9, eaal5272 (2017)
DOI: 10.1126/scitranslmed.aal5272

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Analysis of histone modification patterns in renal tumors.
  • Fig. S2. Histopathological characterization of representative tumor samples.
  • Fig. S3. Genotypic characterization of representative tumor samples.
  • Fig. S4. Molecular characterization of ccRCC lines used in the shRNA minipool screen.
  • Fig. S5. Effects of genetic inactivation of EZH1 or EZH2 on ccRCC.
  • Fig. S6. Characterizing the on-target effects of EZH1 shRNA.
  • Fig. S7. Effects of pharmacological EZH inhibition using JQ-EZ-05 on lymphoid cells.
  • Fig. S8. Effects of pharmacological EZH inhibition using JQ-EZ-05 and GSK126 on renal cells.
  • Fig. S9. Validation of candidate JQ-EZ-05–resistant EZH1 mutants.
  • Fig. S10. Effects of JQ-EZ-05 on VHL−/− cells lacking ARNT1.
  • Fig. S11. Effects of pVHL expression on H3K27 methylation.
  • Fig. S12. Effects of JQ-EZ-05 on H3K27 histone demethylase–deficient VHL−/− ccRCC.
  • Fig. S13. Oncomine analysis of H3K27 demethylases and methyltransferases in renal tumors.
  • Fig. S14. Identification of biomarkers to define EZH1-VHL synthetic lethality.
  • Legends for tables S1 to S7
  • References (5260)

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Other Supplementary Material for this manuscript includes the following:

  • Table S1 (Microsoft Excel format). Annotated list of pLKO-based shRNAs used in the minipool screen.
  • Table S2 (Microsoft Excel format). RIGER analysis to identify genes corresponding to shRNAs that were enriched or depleted selectively in the VHL−/− cells.
  • Table S3 (Microsoft Excel format). RNA-seq data of transcriptional changes observed upon ARNT loss in 786-O cells.
  • Table S4 (Microsoft Excel format). GSEA of 786-O cells with high (VEC) or low (sh/sgARNT) HIF activity.
  • Table S5 (Microsoft Excel format). GSEA of ccRCC cells upon EZH1 inactivation using either pharmacological or genetic tools.
  • Table S6 (Microsoft Excel format). GSEA comparing ccRCC cells that either do (present) or do not (absent) display EZH1-VHL synthetic lethality.
  • Table S7 (Microsoft Excel format). Primer sequences.

[Download Tables S1 to S7]