Figure 7.

cGK inhibition by KT5823 treatment slows down endocytosis. A, Experimental protocol: cells were stimulated with 50 mm KCl for 1 min and the progression of endocytosis was assessed by applying the dye after a variable delay (delay time, Δt). As Δt is prolonged, more endocytosis occurs during this period, reducing the amount of dye subsequently taken up. Dye uptake was quantified by measuring release signals evoked by later KCl stimuli. B, Ensemble averages of measurements of FM l-43 labeled with the fluorescence intensity (a.u.) shown for both control and KT5823-treated cells. C, Ensemble averages of measurements of the FM l-43-labeled showing synaptic vesicle pool size normalized to Δt = 0. The number of individual boutons measured for each condition, where: Δt = 0 s (N = 1105 control, N = 1246 KT 24 h), Δt = 30 s (N = 1816 control, N = 1660 KT 24 h), Δt = 60 s (N = 1200 control, N = 2250 KT 24 h), Δt = 90 s (N = 1216 control, N = 2324 KT 24 h), and Δt = 120 s (N = 893 control, N = 1606 KT 24 h). Data were obtained from n = 3 coverslips obtained from two different cell cultures.