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. 2019 Sep 4;22(9):e25384. doi: 10.1002/jia2.25384

Figure 3.

Figure 3

hCCR5Δ24 mutant has no transdominant negative effect on wtCCR5.(A) HEK‐293T and HeLa‐CD4 cells were transfected with FLAG‐wtCCR5, HA‐hCCRΔ24 and HA‐hCCR5Δ32 alone or cotransfected with FLAG‐wtCCR5 and HA‐hCCRΔ24 or HA‐hCCR5Δ32 in equimolar ratio. (A) GFP reporter vector was added to each transfection in order to analyse CCR5 expression in transfected populations. CCR5 surface or surface + intracellular expression was analysed by flow cytometry using anti‐FLAG and anti‐HA mAbs. (B) Quantification of the flow cytometry experiment from A. (C) HeLa‐CD4 cells were transfected with FLAG‐wtCCR5, HA‐hCCRΔ24 alone or cotransfected with FLAG‐wtCCR5 and HA‐hCCRΔ24 in equimolar ratios. Transfected cells were infected with HXB2, ADA8 or BaL pseudovirus expressing a Luciferase reporter gene. HIV‐1 infection was quantified by measuring Luciferase‐dependent luminescence. Statistical significance was considered when p ≤ 0.05 (****p ≤ 0.0001, ***p ≤ 0.001, **p ≤ 0.01, *p ≤ 0.05; N = 3 independent experiments). Error bars denote mean±SD.