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. 2017 Oct 6;8:1233. doi: 10.3389/fimmu.2017.01233

Figure 4.

Figure 4

C-C chemokine receptor-like 2 (CCRL2) C-terminus is dispensable for receptor internalization. HeLa cells were transfected with a acyl carrier protein (ACP)-CCRL2, or with the truncated forms of ACP-CCRL2 (H8+ or H8−). (A) Total RNA was extracted 24 h after transfection and CCRL2 mRNA expression quantified by qPCR. 2−ΔCT values are reported as average values of two independent experiments performed in triplicate. (B) 24 h after transfection, cell membrane receptors were fluorescently labeled and cells were analyzed by flow cytometry. Data were analyzed using the FlowJo software. The mean values of three independent experiments are shown; median fluorescence intensity of untransfected cells was subtracted and values were normalized over the mean fluorescence intensity of cells expressing non-truncated CCRL2 set at 100. (C) 24 h after transfection, cell membrane ACP-CCRL2 were fluorescently labeled. Cells were then placed at 37°C for 20 min and then fixed. Images were taken using 100× magnification. *p < 0.05 (H8− vs. CCRL2) by Student’s t-test.