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Fig. 1. CCR7 C-terminal deletion mutants reach the plasma membrane. (A) Scheme of CCR7 and its C-terminal deletion mutants. The cytoplasmic C-terminus of CCR7 depicted is based on the prediction by Swiss-Prot (Swiss Institute of Bioinformatics, Basel, Switzerland). Amino acids according to accession number P32248 are indicated. MT1 to MT3 represent C-terminal truncation mutants of CCR7. In the case of MT1, a Lys was replaced by Arg. Potential phosphorylation sites (S, T) and potential ubiquitylation sites (K) are highlighted in bold. The conserved DRY motif in the second intracellular loop was mutated to DNY by site-directed mutagenesis. (B) CCR7 and its C-terminal deletion mutants are equally expressed at the plasma membrane. HEK293 cells stably expressing wild-type (wt) CCR7, MT1, MT2 or MT3 were stained with a FITC-conjugated CCR7-specific antibody and cell surface expression was monitored by flow cytometry. Dashed lines represent untransfected cells stained with the same antibody. FITC-labeled isotype control staining on transfected cells revealed the same results. The experiment has been repeated twice yielding similar results.
