Science Immunology

Supplementary Materials

Supplementary Material for:

Activation of mosquito complement antiplasmodial response requires cellular immunity

Julio César Castillo, Ana Beatriz Barletta Ferreira, Nathanie Trisnadi, Carolina Barillas-Mury*

*Corresponding author. Email: cbarillas{at}niaid.nih.gov

Published 20 January 2017, Sci. Immunol. 2, eaal1505 (2017)
DOI: 10.1126/sciimmunol.aal1505

This PDF file includes:

  • Fig. S1. Confocal imaging of mosquito organs after systemic injection of the lipophilic dye Vybrant CM-DiI.
  • Fig. S2. Confocal staining of midguts of mosquitoes injected with Vybrant CM-DiI, 24 hours after feeding on a P. berghei–infected mouse.
  • Fig. S3. Effect of HDF injection on the percentage of ookinete-invaded cells positive for hemocyte-derived microvesicles (HdMv+).
  • Fig. S4. Effect of HDF injection on P. berghei infection.
  • Fig. S5. Effect of HDF injection on proportion of granulocytes.
  • Fig. S6. Effect of Cactus silencing on the percentage of ookinete-invaded cells positive for hemocyte-derived microvesicles (HdMv+).
  • Fig. S7. Effect of Cactus silencing on P. berghei infection.
  • Fig. S8. Confocal image of polystyrene beads inside a phagocytic granulocyte 24 hours after systemic bead injection.
  • Fig. S9. Effect of hemocyte disruption by systemic injection of polystyrene beads on the percentage of ookinete-invaded cells positive for hemocyte-derived microvesicles (HdMv+).
  • Fig. S10. Effect of hemocyte disruption by bead polystyrene injection on P. berghei infection.
  • Fig. S11. Effect of hemocyte disruption by bead polystyrene injection and infection on the relative expression of several functional markers.
  • Fig. S12. Effect of polystyrene bead injection on the intensity of TEP1 staining on the ookinete surface. immunology.sciencemag.org/cgi/content/full/2/7/eaal1505/DC1
  • Fig. S13. Effect of epithelial nitration on the percentage of ookinete-invaded cells positive for hemocyte-derived microvesicles (HdMv+).
  • Fig. S14. Effect of epithelial nitration on microvesicle release.
  • Fig. S15. Effect of epithelial nitration on the percentage of ookinete-invaded cells positive for hemocyte-derived microvesicles (HdMv+) that are also positive for nitrotyrosine staining.
  • Fig. S16. Top view maximum intensity projection showing the spatial distribution of Sua5.1 cells in direct contact with the surface of a fixed midgut embedded in a collagen matrix.
  • Fig. S17. Viability of Sua5.1 cells associated with the collagen matrix interphase in the absence of physical contact with nitrated or non-nitrated (control) midguts after 6 hours of coincubation.
  • Table S1. Quantification of CM-DiI from dissected midguts.
  • Table S2. Effect of hemocyte disruption by systemic injection of polystyrene beads on the prevalence of Tep1 staining in Plasmodium ookinetes.
  • Table S3. Effect of midgut nitration on microvesicle release.
  • Table S4. Induction of apoptosis (caspase activity) in Sua5.1 cells in response to contact with a nitrated collagen matrix surface for 3 hours.
  • Table S5. Induction of apoptosis (loss of cellular integrity) in Sua5.1 cells in response to 6 hours of exposure to a nitrated midgut and control (non-nitrated).
  • Table S6. List of primers used for gene expression analysis and for dsRNA synthesis.
  • Legends for movies S1 to S5 References (25, 26)

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Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.mov format). Example of the response of Sua5.1 cells to contact with a non-nitrated midgut (replicate #1).
  • Movie S2 (.mov format). Example of the response of Sua5.1 cells to contact with a non-nitrated midgut (replicate #2).
  • Movie S3 (.mov format). Example of the response of Sua5.1 cells to contact with a nitrated midgut (replicate #1).
  • Movie S4 (.mov format). Example of the response of Sua5.1 cells to contact with a nitrated midgut (replicate #2).
  • Movie S5 Movie S5 (.mov format). Example of the response of Sua5.1 cells that are in the same chamber as nitrated midguts but do not come in contact with the midgut surface.

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