Article Figures & Data
Figures
Additional Files
Supplemental data
Files in this Data Supplement:
- supplemental material - QuickTime Movie 1 Time-lapse movie corresponding to Fig. 1 showing the capture of a filament moving through a naturally occurring gap. The images were acquired at 4 second intervals. Note that several focus adjustments were necessary during this movie to compensate for focus drift before and during the flow of perfusate. The white flash designates the point at which the perfusion was initiated. This filament stopped in the gap within four seconds (one time-lapse interval).
- supplemental material - Fig. S1. GFP-NFM incorporation along single neurofilaments. (A) Axonal neurofilaments were induced to splay apart from each other by permeabilization of cultured neurons with 0.5% Triton X-100 in the presence of 0.19M NaCl 3 days after they were transfected by nuclear injection with the GFP-NFM expression vector. Neurofilaments were detected by immunofluorescence microscopy using a polyclonal antibody specific for NFL, which is present in all neurofilaments, and GFP-NFM was detected directly using the intrinsic GFP fluorescence. GFP-NFM incorporation was continuous along some filaments (left) and discontinuous along others (right). The white arrowheads mark the splayed filaments. Scale bar=1�m. (B) Histogram of the proportion of the length of each filament that contained GFP-NFM for 196 filaments from 14 different cells.
- supplemental material - Fig. S2. A captured filament immunostained for NFL. A captured filament, fixed and immunostained using a polyclonal antibody specific for NFL. (A) The intrinsic GFP fluorescence, visualized without an antibody. The image is grainy because the GFP fluorescence is partly quenched during the fixation. (B) The NFL immunofluorescence. The white arrowheads mark the location of the ends of the filament based on the NFL immunofluorescence. Comparison of the length of the captured filament in A and B indicates that this filament had incorporated GFP-NFM along 65% of its length. Scale bar=5�m.