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

letter

. 2014 Oct 20;3(10):e122. doi: 10.1038/oncsis.2014.37

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

Copyright © 2014 Macmillan Publishers Limited

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/

PMC Copyright notice

Chaetocin activity is mediated through ROS and reduces leukemic engraftment. (a) The effect of chaetocin, imatinib and/or BMSFs on the viability of CML-LSCs obtained from the BCR-ABL+, NUP98/HoxA9+ murine model of CML. Cells were pretreated with BMSFs for 4 h and then imatinib and/or chaetocin was added at the indicated concentrations. Viability was measured using trypan blue exclusion 48 h after drug addition and is presented as the percentage of viable cells. (b) The effect of chaetocin, imatinib and/or BMSFs on the induction of apoptosis in CML-LSCs. Cells were treated with imatinib, chaetocin and BMSFs as in panel A. Cells undergoing apoptosis were detected using annexin V FITC and propidium iodide stain following 48 h of drug exposure. The frequency of apoptosis is expressed as the percentage of annexin V positive cells. (c) The effect of imatinib, chaetocin and/or BMSFs on the colony formation of CML-LSCs. Cells were pre-exposed to BMSF for 4 h, treated with imatinib and/or chaetocin for 48 h and plated in methylcellulose at a density of 1000 cells per 150 μl. Colonies were counted after 7 days. (d) Reactive Oxygen Species (ROS) levels in chaetocin-treated cells. CML-LSCs were treated with BMSFs for 4 h and then chaetocin was added at 500 nM. 4 hours after the addition of chaetocin, CellROX Orange was added to achieve 5 μM final concentration. Fluorescence intensity (n=5) is displayed in arbitrary units. (e) The effect of the antioxidant N-acetyl-l-cysteine (NAC) on chaetocin activity. CML-LSCs were pretreated with NAC for 30 min and then treated with BMSFs for 4 h. Following this, chaetocin was added and viability measured 24 h later by trypan staining. (f) The effect of NAC and BMSF on phenethyl isothiocyanate (PEITC) activity. CML-LSCs were pretreated with NAC for 30 min and then treated with BMSFs for 4 h. Following this, PEITC was added and viability measured 24 h later by trypan staining. (g) The effect of chaetocin and BMSFs on leukemia engraftment. CML-LSCs were pre-exposed to BMSF for 4 h, treated with 250 nM chaetocin for 24 h and then injected intravenously into sublethally irradiated mice. Animals were monitored and euthanized when disease symptoms were observed. Five mice were used per treatment condition. P<0.05 chaetocin vs chaetocin/BMSF. Statistical analyses: unpaired t-test (n=3), *P<0.05, **P<0.005 and ***P<0.0005. Error bars represent s.d. from three independent experiments.