Science Immunology

Supplementary Materials

Supplementary Material for:

A PIP2-derived amplification loop fuels the sustained initiation of B cell activation

Chenguang Xu, Hengyi Xie, Xingdong Guo, Haipeng Gong, Lei Liu, Hai Qi, Chenqi Xu, Wanli Liu*

*Corresponding author. Email: liulab{at}tsinghua.edu.cn

Published 17 November 2017, Sci. Immunol. 2, eaan0787 (2017)
DOI: 10.1126/sciimmunol.aan0787

This PDF file includes:

  • Materials and Methods
  • Fig. S1. PIP2 is enriched in the immunological synapse upon B cell activation.
  • Fig. S2. Expression of PIP5Kα, PIP5Kβ, and PIP5Kγ in DT40 B cells.
  • Fig. S3. DGKζ is essential for converting DAG into PA after B cell activation.
  • Fig. S4. BCR signaling–induced PA generation is converted from the PLC-γ2– derived DAG by DGKζ.
  • Fig. S5. The validation of the PIP2 density manipulation system.
  • Fig. S6. Cold treatment induces the regeneration of excessive amount of PIP2 in B cells expressing PLAID-associated PLC-γ2 mutants.
  • Fig. S7. Model depicting the PIP2-derived amplification loop for the robust initiation of B cell activation.
  • Table S1. Sequence of primers used in reverse transcription polymerase chain reaction for detecting the expression of PIP5K and DGK isoenzymes in DT40 B cells.
  • Legends for movies S1 to S5
  • References (42–48).

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Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.avi format). The depletion of PIP2 inside the BCR microclusters and the enrichment of PIP2 at the region outside the BCR microclusters in DT40-WT B cells.
  • Movie S2 (.avi format). PIP2 is depleted inside the BCR microclusters and enriched at the region outside the BCR microclusters in B1-8 primary B cells on a caged-NP antigen system.
  • Movie S3 (.avi format). The depletion of PIP2 inside the BCR microclusters and the enrichment of PIP2 at the region outside the BCR microclusters are impaired in DT40-PLC-γ2-KO B cells.
  • Movie S4 (.avi format). Enrichment of PIP2 at the region outside the BCR microclusters in a PIP5K-dependent manner.
  • Movie S5 (.avi format). DAG is more often than PIP2 to cross the BCR microcluster border.
  • Raw data Excel file

Download Movies S1–S5

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