Supplementary Materials

Supplementary Material for:

Dengue virus NS1 protein activates cells via Toll-like receptor 4 and disrupts endothelial cell monolayer integrity

Naphak Modhiran, Daniel Watterson, David A. Muller, Adele K. Panetta, David P. Sester, Lidong Liu, David A. Hume, Katryn J. Stacey,* Paul R. Young*

*Corresponding author. E-mail: p.young{at} (P.R.Y.); katryn.stacey{at} (K.J.S.)

Published 9 September 2015, Sci. Transl. Med. 7, 304ra142 (2015)
DOI: 10.1126/scitranslmed.aaa3863

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Purification and characterization of DENV-2 NS1 from S2 cells.
  • Fig. S2. Dengue NS1 activates a mouse macrophage NF-κB reporter cell line.
  • Fig. S3. Polymyxin B can efficiently inhibit LPS in the presence of NS1.
  • Fig. S4. NS1 protein causes the loss of surface CSF1R on murine BMMs.
  • Fig. S5. Recombinant NS1 from other flaviviruses has immunostimulatory activity.
  • Fig. S6. Induction of cytokine mRNAs in C57BL/6 BMMs and human PBMCs in response to S2-derived NS1 protein.
  • Fig. S7. NS1 activates cells via TLR4.
  • Fig. S8. Controls for colocalization of NS1 and TLR4 on adherent PBMCs.
  • Fig. S9. Inhibition of PBMC production of IL-8 in response to NS1 by anti-TLR4 antibody and TLR4 antagonist.
  • Table S1. List of mouse primers for RT-PCR.
  • Table S2. List of human primers for RT-PCR.
  • Legend for movie S1
  • References (57, 58)

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Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.avi format). TLR4 and NS1 colocalization on PBMCs; video of immunofluorescence confocal microscopy Z stack.
  • Primary data tables (Excel file)

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[Download Primary data tables]