FIGURE 5.

Strongly reduced membrane expression of α₂δ‐2_R596P in differentiated cultured hippocampal neurons. (a) Representative examples of primary cultured hippocampal neurons transfected with soluble eGFP together with either HA‐tagged WT (2HA‐α₂δ‐2) or mutated (2HA‐α₂δ‐2_R596P) α₂δ‐2. Anti‐HA live‐cell labeling demonstrates a reduced staining intensity of α₂δ‐2_R596P in the soma, dendrites, and the axon (indicated by an arrow) compared to WT α₂δ‐2. (b) Quantification of the average HA fluorescent intensities in the three compartments shows that the surface expression of mutated α₂δ‐2 is strongly reduced compared to WT α₂δ‐2. Graphs show values for individual cells (dots) and mean ± SEM (lines). All values were normalized to the mean of the WT 2HA‐α₂δ‐2 fluorescence intensity within each culture preparation. Data were obtained from three independent culture preparations; 35 and 45 cells expressing WT or mutated HA‐tagged α₂δ‐2 were analyzed, respectively. Statistics: Unpaired t‐test, soma: T ₍₇₈₎ = 3.4; **p = 0.0011, dendrite: T ₍₇₈₎ = 5.1; ***p < 0.0001, axon: T ₍₇₈₎ = 5.5; ***p < 0.0001. Scale bars, 10 μm.