Supplementary Materials

Supplementary Materials for:

Diacylglycerol kinase ζ limits the polarized recruitment of diacylglycerol-enriched organelles to the immune synapse in T cells

Elena Andrada, María Almena, Julia Sáez de Guinoa, Sara V. Merino-Cortes, Rosa Liébana, Raquel Arcos, Silvia Carrasco, Yolanda R. Carrasco, Isabel Merida*

*Corresponding author. Email: imerida{at}cnb.csic.es

This PDF file includes:

  • Fig. S1. Differences between the dynamics of C1bPKCθ and C1a+bPKCθ are due to the recognition of PS by C1a+bPKCθ.
  • Fig. S2. PIP2 dynamics during IS formation.
  • Fig. S3. Dynasore inhibits transferrin endocytosis but not exocytosis during IS formation.
  • Fig. S4. Jurkat cells establish a functional IS with lipid bilayers that present anti-CD3 antibodies.
  • Fig. S5. Generation of the GFP-C1bPKCθ retroviral construct.
  • Fig. S6. Analysis of MTOC and ERC polarization in DGKζ-deficient CTLs and of the effect of DGKα deficiency on MTOC polarization and PKCζ activation.
  • Legend for movies S1 to S7

[Download PDF]

Technical Details

Format: Adobe Acrobat PDF

Size: 969 KB

Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.mp4 format). Dynamic distribution of GFP fused to the tandem C1a+bPKCθ domain during IS formation.
  • Movie S2 (.mp4 format). Dynamic distribution of GFP fused to the PKCθC1a domain during IS formation.
  • Movie S3 (.mp4 format). Dynamic distribution of GFP-fused to the PKCθC1b domain during IS formation.
  • Movie S4 (.mp4 format). Dynamic distribution of the GFP2xPH-Evect2 and Cherry-C1a+bPKCθ constructs during IS formation.
  • Movie S5 (.mp4 format). Dynamic distribution of Cherry-PLCδ PH domain construct during IS formation.
  • Movie S6 (.mp4 format). Loss of the Cherry-PLCδ PH construct from the IS accompanies GFP-C1bPKCθ accumulation when cotransfected Jurkat cells form contacts with SEE-loaded Raji cells.
  • Movie S7 (.mp4 format). Dynamics of the GFP-C1bPKCθ construct in CTLs in contact with peptide-loaded EL4 cells.

[Download Movies S1 to S7]


Citation: E. Andrada, M. Almena, J. S. de Guinoa, S. V. Merino-Cortes, R. Liébana, R. Arcos, S. Carrasco, Y. R. Carrasco, I. Merida, Diacylglycerol kinase ζ limits the polarized recruitment of diacylglycerol-enriched organelles to the immune synapse in T cells. Sci. Signal. 9, ra127 (2016).

© 2016 American Association for the Advancement of Science