HIV-1 Reverse Transcriptase Promotes Tumor Growth and Metastasis Formation via ROS-Dependent Upregulation of Twist

Figure 3

Derivatives of 4T1luc2 cells expressing variants of consensus HIV-1 FSU_A reverse transcriptase (Table 2) exhibit increased levels of ROS production (a–c) and lipid peroxidation (d). ROS production was measured using fluorescent dye DCFH2-DA and normalized to the signal generated by DAPI staining. Total levels of ROS per cell on day 6 of cell culture; production of ROS is quenched by treatment with antioxidants tocopherol (1 μM) and NAC (5 mM); red asterisk indicates significant differences between 4T1luc2_RT-Ann-1.5 and other RT-expressing variants () (a). Total levels of ROS production per cell on day 14 of cell culture relative to the levels exhibited by the parental 4T1luc2 cells (b). ROS levels on day 14 of cell culture normalized to the amount of RT_A variant (in fg) produced by one cell (c). Level of lipid peroxidation on day 14 of cell culture assessed as the concentration of MDA in the lysate of a single cell normalized to the amount of RT_A variant (in fg) produced by one cell (d). Values represent from two independent assays run in duplicates. ns: not significant; ; ; ; (Kruskal-Wallis, followed by Mann-Whitney tests or ordinary one-way ANOVA followed by unpaired -test).

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