The DNA sequences corresponding to human CCR5 (P51681, UniProt), CCR1 (P322246, UniProt), and CXCR4 (P61073, UniProt) were purchased either as synthetic genes (GeneArt, Thermo Fisher Scientific) or as complementary DNA (cDNA) (Open Biosystems). They were cloned into RUSH plasmids downstream of Str-KDEL_IL2ss-SBP-EGFP or Str-KDEL_IL2ss-SBP-mCherry using Fse I and Pac I restriction enzymes (16). Str-KDEL_TNF-SBP-EGFP and Str-KDEL_TNF-SBP-mCherry plasmids have been described elsewhere (16). The CCR5-CCR1tail and CCR1-CCR5tail chimeras were generated from synthetic genes (GeneArt, Thermo Fisher Scientific) and cloned between Fse I and Pac I restriction sites. Mutations from cysteine to alanine in CCR5 tail were generated either by polymerase chain reaction assembly or by the insertion of small synthetic DNA fragments (gBlock from Integrated DNA Technologies). Protein sequences of chimeras and mutants are depicted in Fig. 4B. GFP-CCR5 and GFP-CCR5-Cys3A bear the interleukin-2 (IL-2) signal peptide upstream of GFP and either CCR5 wild-type or CCR5-Cys3A downstream of GFP. A modified version of pEGFP (Clontech) was used for their generation. All plasmids used in this study were verified by sequencing. HeLa cells were transfected using calcium phosphate as described previously (41).

The HIV-1ADA provirus plasmid (pHIV-1ADA) expressing the env gene of the HIV-1 R5-tropic strain ADA has been described elsewhere (42). pNL4.3Δ pNL4.3f was a gift from P. Benaroch (Institut Curie, Paris, France). The plasmid expressing the env gene of VSVG (pEnvVSVG) was a gift from S. Benichou (Institut Cochin, Paris, France).

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