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. 2021 Jan 19;220(3):e201908108. doi: 10.1083/jcb.201908108

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© 2021 Allam et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure S3. — Inhibition of Notch and CXCR4 for 3 h disrupts immunological synapse formation in vitro. Purified DN3a cells were incubated with OP9-DL1 cells for 3 h in the presence or absence of either Notch inhibitor (DBZ) or CXCR4 inhibitor (AMD3100) and then fixed and stained for α-tubulin to mark the MTOC and pTα chain as a marker of pre-TCR (as shown). After triaging for cells in which the MTOC was recruited to the interface (white asterisks) with an OP9-DL1 cell, the percentage of cells in which pTα was polarized to the interface with the OP9-DL1 cell was determined by blind scoring (as shown by column-bar plot). The total number of scored conjugates per condition is 50 (25 cells per biological replicate). Scale bar, 5 µm.