Fig. 2

Rasal1 translocates to the plasma membrane in response to Ca²⁺ in hippocampal neurons. A1-3 Hippocampal neuron expressing Rasal1-EGFP stained to show the distribution of Rasal1-EGFP (green) compared to total Rasal1 immunoreactivity (red) and MAP2 (blue) under basal conditions. Scale bar: 10 µm. B1-3 Depolarized hippocampal neuron expressing Rasal1-EGFP stained to show Rasal1-EGFP (green) distribution compared to total Rasal1 (red) and MAP2 (blue). C Live confocal imaging of neuron expressing Rasal1-EGFP following perfusion of depolarizing buffer (Depolarizing B) C1-4. Followed by perfusion with basal buffer (Basal B) at 8 s (C5-7) followed by second perfusion with depolarizing buffer at 18 s (D-18s) (C8). Scale bar: 10 µm. D–G Rasal1-EGFP live imaging in two additional neurons perfused in basal buffer D, F and depolarizing buffer (E, G). Scale bar: 10 µm. H Time-dependent change in fluorescence ratio (R) in the membrane after addition of depolarizing buffer, basal buffer at 8 s and depolarizing buffer again at 12 s (addition of buffers indicated by dashed lines). Data presented as percentages of the original fluorescence at 0 s. Mean ± SEM are plotted, n = 20