Supplementary Materials

Supplementary Materials for:

Phosphoproteomic analysis of interacting tumor and endothelial cells identifies regulatory mechanisms of transendothelial migration

Marie Locard-Paulet, Lindsay Lim, Giulia Veluscek, Kelly McMahon, John Sinclair, Antoinette van Weverwijk, Jonathan D. Worboys, Yinyin Yuan, Clare M. Isacke, Claus Jørgensen*

*Corresponding author. E-mail: claus.jorgensen{at}cruk.manchester.ac.uk

This PDF file includes:

  • Fig. S1. Comparison of endothelial cell adhesion and transendothelial cell migration of the LM2 and parental MDA-MB-231 cell populations.
  • Fig. S2. Workflow of phosphoproteomic data analysis.
  • Fig. S3. Raw fold changes of the regulated phosphorylation sites identified by MS analysis of tumor cell–endothelial cell coculture.
  • Fig. S4. Raw fold changes of the core set of significantly regulated phosphorylation sites identified by MS analysis of tumor cell–endothelial cell coculture.
  • Fig. S5. Global analysis of quantitative changes occurring at the protein level in tumor cell–endothelial cell cocultures.
  • Fig. S6. Functional validation of EPHA2-knockdown experiments.
  • Fig. S7. Abundance of EPHA2 in parental MDA-MB-231 and LM2 cells and ephrinA1 in HUVECs.
  • Fig. S8. Abundance of overexpressed EPHA2 and EPHA2 mutants in breast cancer cell lines.
  • Fig. S9. Time course of EPHA2 phosphorylation in response to ephrinA1 stimulation of breast cancer cell lines.
  • Fig. S10. Confirmation of an interaction between LMW-PTP and EPHA2.
  • Table S1. Log2(H/M) ratios for regulated phosphorylation sites identified in the LM2 cells by MS analysis of tumor cell–endothelial cell coculture.
  • Table S2. Log2(H/M) ratios for regulated phosphorylation sites identified in the HUVECs by MS analysis of tumor cell–endothelial cell coculture.
  • Table S3. Log2(H/M) ratios for total protein quantification of phosphorylation-regulated proteins in LM2 cells after tumor cell–endothelial cell coculture.
  • Table S4. Log2(H/M) ratios for total protein quantification of phosphorylation-regulated proteins in HUVECs after tumor cell–endothelial cell coculture.
  • Table S5. Total EPHA2 quantification in LM2 cells after tumor cell–endothelial cell coculture.
  • Table S6. Individual siRNA sequences used in the study.
  • Table S7. Nontargeting and negative control siRNA used in knockdown experiments.
  • Table S8. SRM transitions monitored for EPHA2 (wild-type and mutant forms) and LMW-PTP for analyzing protein abundance.

[Download PDF]

Technical Details

Format: Adobe Acrobat PDF

Size: 2.00 MB


Citation: M. Locard-Paulet, L. Lim, G. Veluscek, K. McMahon, J. Sinclair, A. van Weverwijk, J. D. Worboys, Y. Yuan, C. M. Isacke, C. Jørgensen, Phosphoproteomic analysis of interacting tumor and endothelial cells identifies regulatory mechanisms of transendothelial migration. Sci. Signal. 9, ra15 (2016).

© 2016 American Association for the Advancement of Science