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

Figure 5. Activation of Gβγ with mSIRK resulted in an inhibition of DAT activity in heterologous systems.

Figure 5

(A) Oocytes expressing DAT (white bars) or EAAT1 (gray bars) were incubated for 30 min with 0.5% DMSO (DAT/n = 3; EAAT1/n = 25), 5 µM mSIRK (DAT/n = 20, EAAT1/n = 23), or 25 µM mSIRK (DAT/n = 15, EAAT1/n = 15) before uptake assays with [H3]-DA or [3H]-Glutamate, respectively. (B) Dose-dependent inhibition of [3H]-DA uptake in HEK293-DAT cells by mSIRK (n = 4, white circle). (C) 5 min pre-incubation of HEK293-DAT cells with 10 µM mSIRK produced a reduction of Vmax with no changes in Km (n = 3). (D) In HEK293-DAT, 10 µM mSIRK reduced uptake (control, black bar, n = 32; mSIRK, white bar, n = 15). The inhibitory effect of 10 µM mSIRK was attenuated in HEK-DAT cells expressing GRK2ct (control, n = 16; mSIRK, n = 16). **p<0.01 or *p<0.05.