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 Oct 13;10:17095. doi: 10.1038/s41598-020-74105-w

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

© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

In vivo conjugation of GFP-ST/SC to tHBcAg-SC/ST VLPs in the endoplasmic reticulum (ER): The table in the top left indicates the approximate expected sizes of conjugated and unconjugated proteins. Bottom left: Fluorescence imaging of 25 and 70% double sucrose cushion. pEAQ-HT = empty vector control. Unconjugated GFP-ST-ER was mainly detected in the supernatant with some fluorescence in the interface between the 25 and 70% fraction. In presence of tHBcAg-SC-ER VLPs fluorescence was mainly detected in the 70% sucrose fraction indicating conjugation of VLPs and GFP-ST-ER. In contrast tHBcAg-ST-ER and GFP-SC-ER did not seem to result in conjugated VLPs. Right: Western blot (anti HBcAg = top, anti GFP = bottom). Arrow 1 = approximate size of conjugated tHBcAg-ST/SC-ER and GFP-ST/SC-ER (~ 80 kDa), arrow 4 = unconjugated GFP-SC-ER (40 kDa), arrow 5 unconjugated GFP-ST-ER (28 kDa). Unconjugated GFP-ST/SC-ER was mainly detected in the supernatant while tHBcAg-ER was detected in the 70% sucrose fraction. Presence of ~ 80 kDa bands (arrow 1) in 70% fraction in both Western blots indicated successful conjugation of GFP and VLPs. No unconjugated tHBcAg-ST/SC-ER was detected. GFP-SC-ER seems to be mostly unconjugated. Negative control shows that all bands are specific for HBcAg or GFP respectively. Blots cropped for clarity, for uncropped blots see Supplementary Fig. S3.