Tiruchinapalli, et al (2003): Figure 4

Supplementary Material From Singer Lab Publications:

Activity-Dependent Trafficking and Dynamic Localization of Zipcode Binding Protein 1 and beta-Actin mRNA in Dendrites and Spines of Hippocampal Neurons

J Neurosci 23(8):3251-3261 (2003 April 15)

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Figure 4. Rapid stimulation of EGFP-ZBP1 granule transport into dendrites visualized in live neurons after KCl depolarization. Neurons were transfected with EGFP-ZBP1 and imaged live before and after perfusion of 20 mM KCl containing culture medium. A short time lapse (10 frames) was acquired at t = 0, 5, 15, and 30 min after perfusion of KCl.

A, EGFP-ZBP1 signal in proximal dendrite (arrow) from frame 1. Intensity was thresholded so no signal was apparent at distal sites (arrowhead). Fluorescence intensity was displayed as a heat-map (inset).

A', Frame 10 just before KCl perfusion shows comparable signal to Frame 1.

B, At 5 min after KCl perfusion there is a marked increase in EGFP-ZBP1 levels at proximal site (arrow; note warmer colors via the heat map). Some weak signal is apparent at a distal site (arrowhead).

C, D, EGFP-ZBP1 signals continue to increase at 15 and 30 min time points.

E, Plot of timed average fluorescence intensities over time for proximal, middle, and distal dendritic regions of the cell imaged in A-D. Each of the four time lapses is shown in a different color (t = 0, 5, 15, and 30 min).

F, The effects of KCl perfusion-induced increase in ZBP1 localization were analyzed in three live neurons, and the average fluorescence intensities in each region were plotted over time in KCl.