Extended Data Fig. 2: 3[H]-GABA uptake and Inhibitor binding assays.
From: Cryo-EM structure of GABA transporter 1 reveals substrate recognition and transport mechanism
a, Expression profiles of rGAT1WT, rGAT1Epi4 and rGAT1EM observed using FSEC. b, Inhibition of 3H-GABA uptake through rGAT1WT by tiagabine and NO711 displays IC50 values of 918 nM and 563 nM, respectively. Each data point in the graph represents an average of six measurements obtained from two independent sets (n = 2) each done in triplicate with error bars representing SEM (Supplementary Table 1, Supplementary Fig. 3). c, MS-based binding measurements of rGAT1WT expressing membranes with inhibitor NO711 displaying a Kd value of 50 nM. Data points represent specific binding (means ± s.d) calculated from total and non-specific binding measured by LC-MS shown in Supplementary Figs. 4 & 7 of one of three independent experiments (n = 3), each performed in triplicate. Error bars in binding curve represent s.d. d, 3H-GABA uptake of rGAT1Epi4 mutant displaying KM of 3.2 μM (Supplementary Table 1). Each data point in the uptake curve is an average of six measurements obtained from two independent sets (n = 2) each done in triplicate (Supplementary Fig. 5). The error bars represent SEM.
