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. 2015 May 14;134(3):405–415. doi: 10.1111/jnc.13132

Figure 4.

Figure 4

Tetanus toxin (TnTx) abolishes activity‐dependent bulk endocytosis (ADBE). Cultures were preincubated either with or without 2 nM TnTx for 24 h. (a) Cultures were loaded with tetramethylrhodamine‐dextran (TMR‐dextran) (50 μM) during stimulation with a train of 800 action potentials (80 Hz). Quantification of the number of evoked TMR‐dextran puncta per field of view in cultures treated with (green bars) or without (blue bars) TnTx ± SEM. **p = 0.008, Student's t‐test, n = 5 coverslips per condition. (b–e) Cultures were loaded with 10 mM horse radish peroxidase (HRP) during stimulation with a train of 800 action potentials (80 Hz). (b and c) Representative images of nerve terminals after HRP loading in cultures either treated with (+) or without (−) TnTx. Scale bar indicates 200 nm. White arrow indicates HRP‐endosomes, red arrow indicates HRP‐SVs. (c and d) Quantification of the number of HRP‐labelled endosomes (d) or HRP‐labelled SVs (e) ± SEM per nerve terminal in untreated (blue bars) or treated (green bars) cultures. Student's t‐test, ***p < 0.0001, n = 3 independent experiments per condition.