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

. 2020 Jan 1;31(1):27–44. doi: 10.1091/mbc.E19-09-0487

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

© 2020 Tan, Fourriere, et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License.

PMC Copyright notice

FIGURE 3: — Steady-state distribution of BACE1 in HeLa cells and primary neurons. (A) HeLa cells stably expressing BACE1-GFP were fixed, permeabilized, and blocked, followed by staining with either rabbit anti-GCC88 (far-red converted to red/green) and mouse anti-Rab11 (red), mouse anti-EEA1 (red), or mouse anti-CD63 (red) antibodies and DAPI (blue). Insets and higher magnifications of the images of the cells marked * are also shown. Bars represents 10 μm. (B) The percentage of BACE1-GFP at the TGN, recycling endosomes, early endosomes, and late endosomes/lysosomes was calculated as the percentage of total BACE1-GFP pixels that overlapped with GCC88, Rab11, EEA1, and CD63, respectively, using the OBCOL plug-in on ImageJ. Data are represented as the mean ± SD of three independent experiments (n = 16). (C) Primary mouse cortical neurons at DIV7 were stained overnight with anti-rabbit BACE1 for endogenous BACE1 (green) and either human anti-p230/golgin-245 (red), rat anti-LAMP1 (red), mouse anti-Rab11 (red), or anti-Rab4 (red) antibodies at 4°C. Higher magnifications of the merged images are also shown. Bar represents 10 μm. (D) The percentage of endogenous BACE1 at the TGN, recycling endosomes, early endosomes, and late endosomes/lysosomes in neurons was calculated as the percentage of volume sum of BACE1 that overlapped with p230/golgin-245, Rab11, Rab4, and LAMP1, respectively, using Imaris. Data are represented as the mean ± SD of three independent experiments (n = 13–14).