Supplementary Materials

Supplementary Materials for:

mMAPS: A Flow-Proteometric Technique to Analyze Protein-Protein Interactions in Individual Signaling Complexes

Chao-Kai Chou, Heng-Huan Lee, Pei-Hsiang Tsou, Chun-Te Chen, Jung-Mao Hsu, Hirohito Yamaguchi, Ying-Nai Wang, Hong-Jen Lee, Jennifer L. Hsu, Jin-Fong Lee, Jun Kameoka,* Mien-Chie Hung*

*Corresponding author. E-mail: mhung@mdanderson.org (M.-C.H.); kameoka@mail.ece.tamu.edu (J.K.)

This PDF file includes:

  • Fig. S1. Workflow for directly detecting exogenous protein complexes in lysates.
  • Fig. S2. 2D plots of the repeated mMAPS analysis of EGFA647 and EGFRGFP interaction.
  • Fig. S3. Photon burst profile of genomic DNA harvested from cell lysate.
  • Fig. S4. 2D plots of the repeated mMAPS analysis of individual p53GFP and genomic DNA interaction.
  • Fig. S5. 2D plots of the repeated mMAPS analysis of individual p53GFP with either Ser15 phosphorylation, Lys372 methylation, or the control IgG.
  • Fig. S6. Workflow for directly detecting immobilized endogenous proteins in lysates or tissue.
  • Fig. S7. mMAPS analysis for testing the recognition of EGFRGFP by the D38 EGFR antibody.
  • Fig. S8. mMAPS analysis for testing the recognition of EGFRGFP by the ab13 EGFR antibody.
  • Fig. S9. mMAPS analysis for testing the recognition of EGFRGFP by a Flag antibody as a negative control.
  • Fig. S10. mMAPS analysis for testing the recognition of STAT3GFP by a STAT3 antibody.
  • Fig. S11. 2D plots of the repeated mMAPS analysis of endogenous EGFR and EGFA647 interaction with or without EGFR-pTyr1068.
  • Fig. S12. Images and repeats of mMAPS analysis of endogenous EGFR and STAT3 interactions in MDA-MB-468 mouse tumor xenograft tissue.
  • Fig. S13. EGFR and STAT3 interaction ratio from tumor tissues of MDA-MB-468 breast cancer xenograft after antibody efficiency normalization.
  • Fig. S14. Images and 3D plots of the repeated mMAPS analysis of endogenous STAT3, p300, and genomic DNA interactions in MDA-MB-468 mouse tumor xenograft tissue.
  • Fig. S15. Photon burst profiles of specific fluorophores detected by mMAPS platform.
  • Fig. S16. Determination of antibody labeling efficiency for mMAPS.
  • Table S1. Data from each independent experiment used to quantify the ratios of complex formation presented in the figures.

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Citation: C.-K. Chou, H.-H. Lee, P.-H. Tsou, C.-T. Chen, J.-M. Hsu, H. Yamaguchi, Y.-N. Wang, H.-J. Lee, J. L. Hsu, J.-F. Lee, J. Kameoka, M.-C. Hung, mMAPS: A Flow-Proteometric Technique to Analyze Protein-Protein Interactions in Individual Signaling Complexes. Sci. Signal. 7, rs1 (2014).

© 2014 American Association for the Advancement of Science