Figure 5. DNM1a^Ftfl is defective in higher order homo-oligomerization.

(A) Left panel, protein extract from P14 whole brain tissue of homozygous fitful and wildtype littermates incubated with 0 or 20mM EDC cross-linker and hybridized with anti-dynamin-1 antibody. Monomers migrate at 100kD, dimers at 200kD and the tetramers are at 400kD. This assay was performed over three separate times with different samples each time; a representative blot with corresponding percentages is shown. Mean densities (± 1SD) from all experiments are: wildtype 28.75±8.24 (monomer), 29.67±13.9 (dimer), 43.9±8.5 (tetramer); mutant 44±10.7 (monomer), 39.5±12.2 (dimer), 23±13.5 (tetramer) Right panel, COS-7 cells transfected with DNM1-GFP constructs show differences in dimerization. (B) COS-7 cells doubly transfected with DNM1-GFP and DNM1-HA constructs show isoform heterodimerization. Protein extracts from cells were incubated with 0 or 20mM EDC and analyzed by Western blot. Blots were hybridized with anti-GFP antibody, stripped of antibody and then re-hybridized with anti-HA antibody in order to ascertain the presence of each construct in the dimers. A representative blot hybridized with anti-GFP antibody is shown.