Figure 6

Functional effects of miR-510 mediated negative regulation of PRDX1. Western blot of PRDX1 protein expression (A), quantitative real time PCR of peroxiredoxin 1 (PRDX1) mRNA levels (B) and quantified Pacman haptokinetic migration track assays (C) of miR-510 (510) or scrambled control (scr) MCF10A cells transiently transfected with PRDX1 ORF (PRDX1; light gray bars in B) or empty vector (EV; dark gray bars in B). Western blots are normalized to ACTIN and real time PCR to GAPDH. A low and high exposure of the same PRDX1 blot is shown as indicated. Quantitation of haptokinetic migration track assays is of 10 independent tracks per experiment. (D) Transwell migration assay of MCF7 cells stably infected with miR-510 or scrambled (scr) control transiently transfected with PRDX1 or empty vector (EV). (E) Trypan blue exclusion assay of miR-510 expressing (light gray bars) or scrambled control (dark gray bars) MCF10A cells treated with increasing concentrations of hydrogen peroxide (H₂O₂) for 24 h. (F) Trypan blue exclusion assay of miR-510 expressing or scrambled control (scr) MDA MB 231 cells transiently transfected with shPRDX1 vector or empty vector (EV) control for 48 h before treatment with 0 (dark gray bars) and 50 μM (light gray bars) H₂O₂ for 24 h. Data in E and F are plotted as percent live cells compared to untreated control. (E) *P < 0.05 compared to scr control. (F) *P < 0.05 compared to untreated control.