Phosphate buffered salts

Penicillin/streptomycin (Cat # 15140‐122), phosphate‐buffered salts (21300‐058) and Minimal Essential Medium (Cat # 21090‐022) were purchased from Invitrogen (Paisley, Scotland, UK). Foetal bovine serum (Cat # FB‐1001/500, lot # 013BS715) was from Biosera (Nuaille, France). KCl (Cat # BP366) was from Fluka UK (Gillingham, UK). Peptides were synthesized by Genemed Synthesis (San Antonio, TX, USA). The peptides used in this study were as follows – DynIxb_842–851 – GVPRITISDP; DynIxb_842–851AAA –GVARATASDP; DynI_769–784AA – PAGRRAPTSAPTPQRR; DynI_769–784EE – PAGRREPTSEPTPQRR; SydI_356–381 – EWSDDESGNPFGGNEANGGANPFEDD; SydI_356–381AAA – EWSDDESGAAAGGNEANGGAAAAEDD. All peptides were fused to the sequence – RRMKWKK – that permits entry into cells (Lindgren et al. 2000; Cousin et al. 2003) and reconstituted in distilled water. Glutaraldehyde (Cat # R1020) and osmium tetroxide (Cat # AGR1022) were from Agar Scientific (Essex, UK). All other reagents were from Sigma (Poole, UK).

FM1‐43, tetramethylrhodamine‐dextran (TMR‐dextran), penicillin/streptomycin, phosphate‐buffered salts, and Minimal Essential Medium were obtained from Invitrogen. Foetal calf serum was from Biosera. The dynamin I phospho‐specific Ser‐774 antibody was from AbD Serotec, the synaptophysin antibody was from Synaptic Systems and the synaptobrevin II antibody was from Nventa Biopharmaceuticals. Glutaraldehyde and osmium tetroxide were from Agar Scientific. Tetanus toxin (TnTx), BAPTA‐AM (1,2‐bis(o‐aminophenoxy)ethane‐N,N,N′,N′‐tetraacetic acid) and EGTA‐AM were from Calbiochem. Fura‐2 was from Anaspec. Synaptophysin‐pHluorin (sypHy) was a gift from Prof. Leon Lagnado (University of Sussex). All other reagents were from Sigma.