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. 2008 Oct;57(10):2626–2636. doi: 10.2337/db07-1579

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Copyright © 2008, American Diabetes Association

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.

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FIG. 7. — Effect of high glucose (HG) concentration on the production of ROS in MCT cells. A: DCF fluorescence reflecting the relative levels of ROS was imaged with a confocal laser scanning fluorescence microscope in serum-deprived MCT cells treated with high glucose for the indicated time periods. B: DCF fluorescence was measured using the peroxide-sensitive fluorescent probe DCF-DA by a multiwell fluorescence plate reader in intact MCT cells treated with high glucose. Histogram represents means ± SE of three independent experiments. Significant difference from nontreated cells is indicated by *P < 0.05 and **P < 0.01. C: NAC blocks ROS generation measured using the peroxide-sensitive fluorescent probe DCF-DA in MCT cells treated with high glucose. Histogram represents means ± SE of three independent experiments. Significant difference from normal glucose (NG) is indicated by **P < 0.01 and from cells treated with NAC versus high glucose alone is indicated by ††P < 0.01. D: Effect of NAC on high glucose–induced Akt/PKB and tuberin phosphorylation and OGG1 expression in MCT cells. Representative immunoblot shows a decrease in phospho-Akt (p-Akt) and phospho-tuberin (p-tuberin) and an increase in OGG1 expression in cells preincubated with NAC before exposure to high glucose. E: NAC blocks 8-oxodG generation in cells treated with high glucose. Immunoflourescence shows a decrease in mitochondrial and nuclear 8-oxodG staining in MCT pretreated with NAC compared with cells treated with high glucose alone for 2 h. (Please see http://dx.doi.org/10.2337/db07-1579 for a high-quality digital representation of this image.)

FIG. 7. — Effect of high glucose (HG) concentration on the production of ROS in MCT cells. A: DCF fluorescence reflecting the relative levels of ROS was imaged with a confocal laser scanning fluorescence microscope in serum-deprived MCT cells treated with high glucose for the indicated time periods. B: DCF fluorescence was measured using the peroxide-sensitive fluorescent probe DCF-DA by a multiwell fluorescence plate reader in intact MCT cells treated with high glucose. Histogram represents means ± SE of three independent experiments. Significant difference from nontreated cells is indicated by *P < 0.05 and **P < 0.01. C: NAC blocks ROS generation measured using the peroxide-sensitive fluorescent probe DCF-DA in MCT cells treated with high glucose. Histogram represents means ± SE of three independent experiments. Significant difference from normal glucose (NG) is indicated by **P < 0.01 and from cells treated with NAC versus high glucose alone is indicated by ††P < 0.01. D: Effect of NAC on high glucose–induced Akt/PKB and tuberin phosphorylation and OGG1 expression in MCT cells. Representative immunoblot shows a decrease in phospho-Akt (p-Akt) and phospho-tuberin (p-tuberin) and an increase in OGG1 expression in cells preincubated with NAC before exposure to high glucose. E: NAC blocks 8-oxodG generation in cells treated with high glucose. Immunoflourescence shows a decrease in mitochondrial and nuclear 8-oxodG staining in MCT pretreated with NAC compared with cells treated with high glucose alone for 2 h. (Please see http://dx.doi.org/10.2337/db07-1579 for a high-quality digital representation of this image.)