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. 2010 Oct 6;20(1):80–89. doi: 10.1093/hmg/ddq434

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© The Author 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — PTEN ATP-binding mutations increase ROS production and SOD1 expression. (A) MCF-7 cells stably expressing empty vector, PTEN^WT or PTEN ATP binding mutants were incubated with 25 μm Carboxy-H2DCFDA for 30 min and immunofluorescent images were taken for ROS production (green). DNA was counterstained with Hoechst 33342 (blue). (B) Quantitation of immunofluorescence intensity of DCF [from (A)] in MCF-7 cells overexpressing PTEN^WT and mutants. The data represent the average of three independent measurements. P < 0.01, one-way ANOVA. (C) SOD1 protein was detected in cell lines expressing WT or mutant PTEN by western blot. (D) Total RNA from MCF-7 cells overexpressing WT or mutant PTEN was extracted and qRT–PCR was performed to measure TP53INP1 mRNA levels. P < 0.01, one-way ANOVA.