Figure 7.

Cellular abundance of Prdx6 was required for significant protection of SRA-hLECs by Bmal1, and reduced abundance of Prdx6 affected the protective potential of Bmal1. SRA-hLECs were transfected with Prdx6-As or empty–vector. After 48 h, cells of each group were pooled and transfected with pGFP-vector or pGFP-Bmal1 using the Neon transfection system (Invitrogen). Equal numbers of cells were harvested in 96-well plates for MTS and ROS assays to avoid the transfection effect and then treated or untreated with different concentrations of H₂O₂ as indicated. (A) Six hours after H₂O₂ exposure, ROS intensity was measured using CellROX Red reagent as described in Materials and Methods. (B) Cell viability was measured after 16 h of H₂O₂ exposure using MTS dye. Data represent means ± S.D. values of three independent experiments. Black bars vs. red bars; *p < 0.05, **p < 0.001. (C) SRA-hLECs were transfected with antisense (As)-Prdx6 (4 µg) or empty vector plasmid, and the effect of antisense of Prdx6 (As-Prdx6) was confirmed through immunoblotting with an anti-Prdx6 antibody.