Figure 5. Blocking NSF-dependent de-priming occludes the effect of Syt-7 at low prestimulation [Ca²⁺].

(A,B) Calcium uncaging experiment from low prestimulation [Ca²⁺] in WT (A) and in Syt-7 KO (B) control cells (Control, gray) and in cells infused with 200 µM N-Ethylmaleimide (+NEM, magenta). Panels are arranged as in Figure 1A, except that amperometric measurements were not included. (C) Size of the burst (secretion within 0.5 s after the flash). (D) Total release (secretion within 5 s after the flash). (E) Size of the RRP. (F) Time constant, τ, of fusion for fast (i.e. RRP). (G) Size of the SRP. (H) Time constant, τ, of fusion for slow (i.e. SRP). Data information: Data are presented as mean ± SEM. *: p<0.05; **: p<0.01, Mann Whitney test comparing control cells to cells infused with NEM from the same genotype. Number of cells: WT control: N = 15 cells; WT + NEM: N = 15 cells; Syt-7 KO control: N = 24 cells; Syt-7 KO + NEM: N = 23 cells.

Figure 5—figure supplement 1. Blocking NSF had no effect in WT and Syt-7 KO cells when stimulated from high prestimulation [Ca²⁺].

(A,B) Calcium uncaging experiment from high prestimulation [Ca²⁺] in WT (A) and in Syt-7 KO (B) control cells (Control, gray) and in cells infused with 200 µM N-Ethylmaleimide (+NEM, magenta). Panels are arranged as in Figure 1A, except that amperometric measurements are not included. (C) Size of the burst (secretion within 0.5 s after the flash). (D) Total release (secretion within 5 s after the flash). (E) Size of the RRP. (F) Time constant, τ, of fusion for fast (i.e. RRP). (G) Size of the SRP. (H) Time constant, τ, of fusion for slow (i.e. SRP). Data information: Data are presented as mean ± SEM. *: p<0.05; **: p<0.01, Mann-Whitney test comparing control cells to cells infused with NEM from the same genotype. Number of cells: WT control: N = 20 cells; WT + NEM: N = 24 cells; Syt-7 KO control: N = 15 cells; Syt-7 KO + NEM: N = 17 cells.