Fig. 1 | Scientific Data

Fig. 1

From: High-throughput analysis of the human thymic Vδ1+ T cell receptor repertoire

Fig. 1

Summary of donor details and experimental workflow employed in this study. FACS-sorted CD3+ TCRVδ1+ TCRVδ2 thymocytes were analysed at the mRNA level by next-generation sequencing of CDR3 regions of TRG and TRD. (a) Name, age (d = days, m = months), sex, purity and number of sorted cells of the analysed samples (n = 8). (b,c) Pre- and post-sorting gating strategy. CD3+ TCRαβ TCRVδ1+ TCRVδ2 cells were sorted to > 98% final purity. After selection of Singlets (FSC-A x FSC-W) and exclusion of debris/dead cells (FSC-A x SSC-A), Vδ1+ cells were sorted from the CD3+αβ population. (d) Amplicons were generated from sorted Vδ1+ thymocyte by mRNA/cDNA based multiplex PCR technology. Multiplex primer sets amplify CDR3 regions by targeting Vγ or Vδ and constant gene segments, with the addition of Illumina sequencing adapters as overhangs (red). Sequences were obtained by Illumina MiSeq sequencing and next annotated by IMGT as described in the methods section before downstream bioinformatics analysis. (ef) Hierarchical clustering of thymic and PB TRG and TRD samples using F pairwise similarity metric. Samples were clustered by age (months and days were normalized per year in order to have the same unit of measure - left) and sex (right).

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