Fig. 7
Hydroxy-Tam exposure increases oxidative stress. a MCF7 and MCF7-HTR whole-cell lysates were used for analysis of total MnSOD activity. b, c MCF7 and MCF7-HTR, and T47D and T47D-HTR whole-cell homogenates, were used to determine: b steady-state levels of O2●−, by MitoSox oxidation; and c H2O2 by CDCFH2 oxidation. d Glutathione levels in MCF7 and MCF7-HTR whole-cell homogenates. e Semi-native gel analysis of MCF7 and MCF7-HTR, as well as T47D and T47D-HTR cell lysates. f–h Whole-cell homogenates of MCF7-HTR cells expressing MnSODWT, MnSODK68Q, or MnSODK68R were analyzed for: f steady-state levels of O2●−, g H2O2, and h glutathione levels. i MCF7 and MCF7-HTR cells were stained for Ki-67 and DAPI. j, k Clonogenic survival experiments for MCF7-MnSODK68Q cells expressing AdMitoCat. Cell were treated with 1 μM hydroxy-Tam for 120 h and k quantified. All experiments were done in triplicate. Error bars represent ± 1 SEM. *p < 0.05 and ***p < 0.001. Three groups analyzed via a one-way ANOVA statistical analysis with Tukey’s post-analysis and two groups analyzed by a t-test
