Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Nov 2;13:6558. doi: 10.1038/s41467-022-34052-8

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 5 — Rat hippocampal neurons were activated with TTX withdrawal with or without 250 nM torin-1. a Total cell lysates were analyzed by western blotting with the antibodies indicated next to the corresponding blot (n = 2). b Diffusion heat map of Halo-eIF4E trajectories obtained in inactivated neurons (16 h TTX) at a frame rate of 100 Hz (with a dotted neuronal outline, scale bar: 5 μm. Each point in the image is false‐colored according to the mean square displacement calculated over all displacements originating in a circle (r = 80 nm). Inset: Trajectories and corresponding diffusion heat map (scale bar: 1 μm). c Diffusion map of Halo-eIF4E of activated neurons after TTX withdrawal (scale bar: 5 μm). Inset: Trajectories and corresponding diffusion heat map (scale bar: 1 μm). d Diffusion speed of Halo-eIF4E in neurons treated with 250 nM torin-1 (red colors) (scale bar: 5 μm). e Single particle tracking was recorded at a frame rate of 200 Hz. The cumulative distribution function (CDF) of all single-molecule displacements for Halo-eIF4E is shifted to the left when compared to neurons treated with torin-1, which indicates a shift towards slower diffusion for Halo-eIF4E undergoing cap-dependent translation. CDF was obtained from 1226 and 742 trajectories in 10 individual dendrites of neurons treated with vehicle control or torin-1, respectively. The global two-component fit returns a global apparent D_slow of 0.32 ± 0.006 µm²/s and a global apparent D_fast of 4.10 ± 0.076 µm²/s. While the majority of control Halo-eIF4E exhibit slow diffusion (76 ± 0.6%), this percentage drops to 48 ± 0.6% when treated with torin-1. The data was recorded at 200 Hz, and we report apparent diffusion coefficients for that frame rate. f Left: Medium-intensity projection of 10,000 frames of _JF549Halo-eIF4E recorded simultaneously with sparsely photo-activated _PA-JF646Halo-eIF4E molecules (individual _PA-JF646Halo-eIF4E trajectories are shown in green) at 56 Hz frame rate (scale bar: 10 μm). Right: The associated _PA-JF646Halo-eIF4E diffusion heat map depicts that slow Halo-eIF4E molecules (in blue) linger near what appear to be spines in activated neurons (scale bar: 10 μm). Spines are indicated with an asterisk.