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. 2013 Mar 26;8(3):e59788. doi: 10.1371/journal.pone.0059788

Figure 4. Overexpression of Gβγ subunits results in decreased DAT uptake activity in HEK-293-DAT cells.

Figure 4

(A, B) Specific [3H]-DA uptake and kinetic analysis for HEK293-DAT control cells (white bar, n = 6) or transfected with Gβ1γ2 (n = 3), Gαi2 (n = 3), or Gαi2Gβ1γ2 (n = 3). (C) Overexpression of Gβ4 (n = 5) or Gβ5 (n = 7) (gray bars), decreased [3H]-DA uptake in HEK293-DAT cells when compared to control (white bar, n = 19). Overexpression of the Gβγ scavengers (black bars), Gαt (n = 6), or GRK2ct (n = 4) increased [3H]-DA uptake. (D, E) Overexpression of Gβ1γ2 (n = 4), Gβ4 (n = 5), or Gβ5 (n = 7) in HEK-DAT cells did not alter the plasma membrane levels of DAT as measured by biotinylation. T = total fraction, B = biotinylated fractions, **p<0.01.