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 Oct 27;10(10):e005535. doi: 10.1136/jitc-2022-005535

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

© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

PMC Copyright notice

Lack of cytotoxicity against cells naturally expressing or overexpressing mutant H3.3 histone. (A) 15×10³ transduced CD8⁺ cells were cocultured at a 5:1 E:T ratio with HLA-A*02:01⁺ Mel21a cells, retrovirally transduced to express either H3.3K27M full length cDNA (MP71_H3.3K27M-CDS-P2A-GFP) (top panel) or CDK4R24 full length cDNA (MP71-CDK4-R24L-P2A-GFP) (bottom panel). Cytotoxicity was observed over 72 hours using the live cell imaging system IncuCyte Zoom (Essen bioscience). Representative images are shown in the right panels. Values are calculated by normalizing the average GFP total area (µm²/image) in the target cells cocultured with the respective TCR-transduced T cells to the average of that cocultured with mock transduced T cells. The experiment was performed three times with similar results and graphs represent means of triplicate cultures±SD. (B) 15×10³ CD8⁺ transduced cells were cocultured at a 5:1 E:T ratio with SF8628-A2 cells, naturally expressing the H3.3K27M mutation and additionally either retrovirally transduced to express CDK4R24L full length cDNA (top panel) or loaded with 10^-6 M mutant H3.3K27M peptide (bottom panel). Cytotoxicity was observed over 72 hours using the live cell imaging system IncuCyte Zoom. Representative images are shown in the right panels. Values are calculated as in (A). The experiment was performed three times with similar results and graphs represent means of triplicate cultures±SD. GFP, green fluorescent protein.