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

. 2021 Aug 26;12:5131. doi: 10.1038/s41467-021-25448-z

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

© The Author(s) 2021

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. 1 — a Schematic diagram of different GFP_1–₁₀-NLS-related recombinant proteins. The amino acid sequences of TAT, INF7, NLS, 6 × His and the different proteolytic cleavage sites (N and Na-Nf) are presented below, and each protease-recognised cleavage site is labelled in brackets. b Schematics of the endonuclear split GFP assay. Only after GFP_1–₁₀-NLS was released from the endosome and translocated into the nucleus of HEK-293T cells with stable expression of histone H3-GFP β-sheet 11 (HEK-293T GFP₁₁), the fluorescence of the complete GFP in the nucleus observed, and co-localised with the mRuby3 fluorescence. c The MFI of HEK-293T GFP₁₁ cells treated with GFP_1–₁₀-NLS-related proteins containing different cleavage sites (5 μM). d The MFI (left panel) within total HEK-293T cells and the corresponding percentage of green fluorescence-positive cells (right panel) treated with GFP_1–₁₀-NLS-related proteins containing two cleavage sites (5 μM). e Immunoblot analysis of GFP and f the MFI in the treated cells at different time points during the endonuclear split GFP assay (1 μM). g Representative microscopy images of cytosolic fluorescence distribution in MA-104 cells treated with different GFP-related proteins (1 μM) as indicated (upper panel: scheme diagram of GFP-related constructs). h The MFI (left panel) within total HEK-293T cells and the corresponding percentage of green fluorescence-positive cells (right panel) after treatment with GFP-related proteins (1 μM). For (c, d, f, and h) the results shown are the means ± s.e.m.; n = 3 biologically independent samples; *P < 0.05, **P < 0.01, ***P < 0.001, and NS no significant difference; two-tailed unpaired student’s t test. Relative MFI (fold increase) was obtained by MFI of total cells treated with the indicated proteins divided by that of total cells treated with the corresponding cargo protein only. For (b bottom panel) and (g bottom panel), the data shown are representative of three independent experiments, respectively; for (e), the data shown are representative of two independent experiments. For data, statistics, exact P values and uncropped images of the immunoblots, see Source Data File.