Supplementary Figure 5: Reactivity of H1 and C2 hybridomas.
From: Deciphering CD4+ T cell specificity using novel MHC–TCR chimeric receptors
(a) T cell hybridomas H1 and C2 were stained with TCR Vbeta specific antibodies. All cells showed positive staining only with the TCR Vbeta14 antibody. (b) cDNA from hybridomas H1 and C2 was used to PCR and clone the TCRbeta chains using oligos specific for different TCRV beta chains. For both hybridomas, apart from the nonfunctional TCRb chain from the BW fusion partner (not shown) a functional TCRbeta containing the TCR Vbeta14 and TCRJbeta2–5 was cloned. A comparison of the protein sequences is shown, highlighting the different CDR3 regions. (c) Hybridomas carrying the NFAT-GFP reporter were co-cultured (n = 2 co-cultures) with BMDCs pulsed with different amounts of the indicated peptides (μg/ml, 90% purity). The graph shows NFAT activation (GFP). Data are representative of two independent experiments. (d) Half maximal effective concentration (EC50) was calculated by non-linear regression (curve fit) based on data from two experiments, mean ± sd are shown.
