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

. 2017 Sep 4;8:423. doi: 10.1038/s41467-017-00448-0

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

© The Author(s) 2017

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. 6 — Genetically engineered human RBCs made in culture and expressing GPA-VNA/A or Kell-VNA/A differentiate normally and protect neurons against BoNT/A challenge both in neuronal culture and in vivo. a Mobilized human CD34+ cells infected with lentiviruses expressing the chimeric GPA-VNA/A protein were cultured by the method detailed in Fig. 4 and the expression of multiple surface proteins was examined by flow cytometry at the indicated time points. b Upper panel shows CD235A and Hoechst staining of human cells expressing GPA-VNA/A generated from CD34+ cells that have been cultured in vitro for 20 and 23 days. Lower panel shows Giemsa and hemoglobin staining of hRBCs expressing GPA-VNA/A at d20 and d23. c Proliferation curve during culture of mobilized human CD34+ cells expressing vector or GPA-VNA/A. (n = 3/group, mean ± S.E.M.). d Human RBCs expressing GPA-VNA/A or Kell-VNA/A protect cultured neuronal cells from BoNT/A protease activity. Rat neurons were co-incubated with BoNT/A and engineered hRBCs as indicated and neuronal lysates were analyzed by western blotting as in Fig. 1. e Survival curve of mice challenged with BoNT/A. Human RBCs generated by in vitro culture were submitted to flow cytometry to detect the percentage of GFP+ cells before injecting into mice. One hundred and fifty million GFP+ human RBCs expressing empty vector or GPA-VNA/A were transfused into NOD/SCID mice. After 30 min, the mice were challenged with 10 LD₅₀ BoNT/A and were observed for 7 days (n = 4/group). Mice were challenged with 10 LD₅₀ BoNT/A 1 day after injection of 120 million GFP+ human RBCs and was observed for 7 days (n = 2/group)