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. 2016 Feb 10;6:20508. doi: 10.1038/srep20508

Figure 6. Expression of OCT1-EYFP, but not the OCT1-D474C variant, increases DNR uptake in HEK293T cells.

Figure 6

The plasmid pOCT1-EYFP was used to create the OCT1-D474C mutation by site-directed mutagenesis. The plasmids pEYFP (empty), pOCT1-EYFP and pOCT1-D474C-EYFP were transiently transfected into HEK293T cells and exposed to DNR. Cells expressing EYFP were sorted to quantify the level of DNR uptake using FACS analysis using FL1 530/30 to detect EYFP fluorescence and FL4 670LP for DNR fluorescence. (A) Epifluorescent microscopy showing localization of the OCT1-EYFP fusion protein. Pictures were taken with Ziess Z2 fluorescent microcope using DAPI filter set and EYFP filter set. Results are representative of three separate experiments. (B) DNR uptake in HEK293T cells carrying either the plasmid pOCT1-EYFP, the variant, or the empty EYFP vector. Results are expressed as the mean ± S.D. from three separate experiments. (C) Comparison of the rhodamine uptake level in HEK293T cells carrying the empty EYFP vector, or either the plasmid pOCT1-EYFP or pOCT1-D474C-EYFP. Results are expressed as the mean ± S.D. from three separate experiments.