FIGURE 1.
ROS levels in MEFs. A, prx1+/+ and prx1-/- MEFs were simultaneously stained with 0.25 μm CM-H2-DCFDA and analyzed by flow cytometry. The curves are representative of several independent experiments with similar outcomes. B, mean values ± S.E. of at least five separate experiments from A were calculated as described under “Experimental Procedures.” ROS levels in prx1+/+ cells were arbitrarily set at 1. C, prx1+/+ and prx1-/- MEFs were grown on glass coverslips and then exposed to 2.5 μm CM-H2-DCFDA. Cells were then immediately fixed in paraformaldehyde and imaged by fluorescence microscopy. Images were captured with a ×100 objective. D, prx1-/- cells were transduced with a prx1-expressing lentivirus or a control vector. Immunoblot (inset) shows expression of the protein. β-Tubulin was used as a loading control. Cells were stained with 0.25 μm CM-H2-DCFDA and analyzed by flow cytometry in at least three separate experiments. Mean values ± S.E. of at least five separate experiments were calculated as described under “Experimental Procedures.” ROS levels are expressed relative to that of the prx1+/+ cells from A and B, which was arbitrarily set at 1. E, prx1-/- cells reconstituted with lentiviral prx1 (D) were imaged by fluorescence microscopy, as described for C.