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(A) Single pTα CD4CD8CD25 TCRγδ cells were sorted onto OP9-DL1 monolayers

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posted on 2011-12-31, 17:11 authored by Taras Kreslavsky, Annette I. Garbe, Andreas Krueger, Harald von Boehmer
On day 7, the contents of each well were divided into 3 parts. One part was analyzed by FACS for surface marker expression (CD45, CD4, CD8, TCRγδ, and TCRβ), the other two parts were transferred to wells with fresh OP9-DL1 monolayers coated with TCRγδ antibodies or uncoated. On day 14 cells were again analyzed for surface marker expression, and cells from anti-TCRγδ–coated wells were further transferred to uncoated wells with fresh OP9DL1 monolayers and analyzed for marker expression on day 19. (B) The proportion of wells containing DN or both DN and DP cells is shown. Numbers represent percentages of particular wells among the total number of wells that contained CD45 cells at any time point. (C) A representative well containing γδ lineage cells only is shown. CD4CD8TCRγδ cells are detected at least at one time point, and CD4CD8 cells are absent at all time points. (D) A representative well containing αβ and γδ lineage cells only is shown. CD4CD8 cells are present before anti-TCRγδ stimulation. Only CD4CD8TCRγδ cells are detected in the presence of anti-TCRγδ antibody and after its removal. Histogram below shows comparison of surface expression of TCRγδ on DN and DP cells in the same well on day 14 of culture without antibody. Numbers in quadrants indicate the percentage of cells. Red numbers above plots refer to absolute cell number.

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Taken from "T cell receptor–instructed αβ versus γδ lineage commitment revealed by single-cell analysis"

The Journal of Experimental Medicine 2008;205(5):1173-1186.

Published online 12 May 2008

PMCID:PMC2373848.

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