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. 2010 Jun 15;285(35):27289–27301. doi: 10.1074/jbc.M110.131003

FIGURE 1.

FIGURE 1.

Thirteen-transmembrane segment fusion protein between DAT and Tac contains high affinity FLAG epitope and is functionally expressed in plasma membrane of HEK293 cells. A, TacDAT is a head-to-tail fusion of hDAT (blue) and Tac (green) with an N-terminal M1 antibody FLAG epitope, DYKDDDDK. B, non-permeabilizing immunostaining of HEK293 cells transfected with TacDAT using an M1 antibody recognizing the FLAG epitope. Scale bar, 10 μm. C, [3H]dopamine (DA) uptake experiments in HEK293 cells transfected with TacDAT (green) or hDAT (blue). The data are shown as relative uptake in percentage of [3H]dopamine uptake in wild type DAT (means ± S.E., n = 3 of triplicate determinations). D, [3H]dopamine uptake after preincubating HEK293 cells expressing TacDAT or hDAT for 30 min at 4 °C with M1 antibody. Data are means ± S.E. of n = 3. E, surface HA.11 and M1 ELISA signals from HEK293 cells expressing TacDAT with an inserted HA tag in the second extracellular loop (Tac-HA-DAT). Values are means ± S.E. of n = 3. A.U., arbitrary units.