Figure 3
Functional analysis of α1-GFP GlyR nanodiscs from HEK293 cells and X. laevis oocytes. (a) Dose–response relationship of α1-GFP GlyR obtained by MST experiments and electrophysiological recordings from HEK293 cells and oocytes. MST data points were inversely normalized for better comparison with dose–response curves obtained from electrophysiological measurements. Data are shown in mean ± SEM. (b) Western blot of SMA-copolymer solubilized GFP-GlyR α1 obtained from the membrane fractions of oocytes and HEK293 cells, show a single band at the calculated molecular weight below 70 kDa. Western blot image was cropped, indicated by a grey cropping line. (c) Electrophysiological experiments obtained from oocytes and HEK293 cells revealed aEC50 values 212.9 ± 21 µM and 68.8 ± 7.4 µM, respectively. cEC50 values of 52.6 ± 40.8 µM (n = 4) and 40.9 ± 13.4 µM (n = 4) obtained from oocytes and HEK293 cells showing no significant difference (p = 0.41). The aEC50 obtained from is significantly higher (p < 0.01) than the measured cEC50, while the aEC50 and cEC50 obtained from HEK293 cells show no difference (p = 0.23). Error bars represent SEM between independent experiments. (d) Signal amplitudes obtained from MST experiments reveal no differences between HEK293 cells (signal amplitude = 4.04 ± 1.09) and oocytes (signal amplitude = 3.38 ± 1.09; p = 0.41, n = 4). Data are shown in mean ± SD. Unpaired two-side t test for statistics.
