Supplementary Materials

Supplementary Material for:

Dormant breast cancer micrometastases reside in specific bone marrow niches that regulate their transit to and from bone

Trevor T. Price, Monika L. Burness, Ayelet Sivan, Matthew J. Warner, Renee Cheng, Clara H. Lee, Lindsey Olivere, Karrie Comatas, John Magnani, H. Kim Lyerly, Qing Cheng, Chad M. McCall, Dorothy A. Sipkins*

*Corresponding author. Email: dorothy.sipkins{at}dm.duke.edu

Published 25 May 2016, Sci. Transl. Med. 8, 340ra73 (2016)
DOI: 10.1126/scitranslmed.aad4059

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Bone homing capacity of ER+ and ER BCC lines.
  • Fig. S2. Intracellular CXCR4 stores expressed in ER+ and ER cell lines.
  • Fig. S3. Schematic for CXCR4-mediated homing inhibition experiments.
  • Fig. S4. Effect of CXCR4 siRNA knockdown on BCC BM homing.
  • Fig. S5. E-selectin ligand and synthetic enzyme mRNA transcript expression in ER+ and ER BCC lines and primary BCCs.
  • Fig. S6. E-selectin ligand protein expression in ER+ and ER BCC lines.
  • Fig. S7. Flow cytometric analysis of E-selectin ligand expression on BCC lines.
  • Fig. S8. CD44 cell surface expression on ER+ and ER BCC lines.
  • Fig. S9. Immunofluorescence analysis of E-selectin ligand expression on BCC lines.
  • Fig. S10. Western blot analysis of E-selectin ligand expression profiles in BCC lines.
  • Fig. S11. Western blot analysis of E-selectin ligand expression profiles in BCC lines, with protection of sialic acid residues.
  • Fig. S12. Immunofluorescence analysis of MUC1 expression on ER+ and ER BCCs.
  • Fig. S13. Immunoprecipitation and functional analysis of the E-selectin binding capacity of MUC1 in BCC lines.
  • Fig. S14. Western blot analysis of E-selectin ligand expression profiles on MCF-7 stem and nonstem cell populations.
  • Fig. S15. Schematic for E-selectin–mediated homing inhibition experiments.
  • Fig. S16. Immunofluorescence analysis of E-selectin ligand expression on MCF-7 stem and nonstem cell populations.
  • Fig. S17. Fluorescent lipophilic membrane dye depletion after successive cell doubling events.
  • Fig. S18. Spontaneous metastasis of orthotopically engrafted MDA-MB-231 (DiD+tdT+) BCCs to the calvarial BM.
  • Fig. S19. Schematic for orthotopic engraftment and E-selectin–mediated homing inhibition experiments.
  • Fig. S20. Schematic for GMI-1271 and AMD3100 mobilization experiments.
  • Fig. S21. Schematic for AMD3100 mobilization experiments in mice with established disease.
  • Fig. S22. AMD3100-induced mobilization of dormant BCCs out of the BM and into the circulation.
  • Fig. S23. Lack of effect of GMI-1271 on BCC mobilization from BM.
  • Fig. S24. Montage image of primary human BCC homing to sinusoidal regions of the calvarial BM.
  • Table S1. Gene list.
  • Table S2. Cell lines.
  • Legends for videos S1 to S6
  • Reference (48)

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Other Supplementary Material for this manuscript includes the following:

  • Video S1 (.mov format). Circulating BCCs crawling along vascular wall (merge).
  • Video S2 (.mov format). Circulating BCCs crawling along vascular wall (DiD channel only).
  • Video S3 (.mov format). Circulating BCCs before AMD3100 treatment (merge).
  • Video S4 (.mov format). Circulating BCCs before AMD3100 treatment (DiD channel only).
  • Video S5 (.mov format). Circulating BCCs after AMD3100 treatment (merge).
  • Video S6 (.mov format). Circulating BCCs after AMD3100 treatment (DiD channel only).