Fig. 2. Effect of H₂O₂ in wild type (WT) and Grx2 knockout (Grx2 KO) lens epithelial cells.

(A) Comparison of the cell density and morphology after H₂O₂ treatment. WT and Grx2 Grx2 KO primary mouse LECs were treated with or without 100 μM H₂O₂ for 6 hrs. After treatment, cell morphology was observed under phase contrast microscopy. (B) Effect of H₂O₂ on cell viability in WT and Grx2 KO cells. WT and Grx2 KO primary mouse LECs were plated in 96-well plate (all normalized to 10,000 cells/well) and treated with or without 100 μM H₂O₂ at for indicated times. Cell viability in each time point was measured by adding WST-8 reagent and then the transmission was evaluated under 450 nm using a 96-well microplate reader. Error bars indicate S.D., n=3, *P<0.05 vs. WT treated with H₂O₂. (C) Effect of H₂O₂ on LDH release in WT and Grx2 KO cells. Wild type and Grx2 knockout primary mouse LECs were treated with or without 100 μM H₂O₂ at different time point from 0 to 6 hours. LDH activity was measured as described as under “Materials and methods”. Error bars indicate S.D., n=3, *P<0.05 vs. WT treated with H₂O₂. (D) Effect of H₂O₂ on caspase 3/7 activation in WT and Grx2 KO cells. Wild type and Grx2 knockout primary mouse LECs were plated in 96-well plate (all normalized to 10,000 cells/well) and treated with 0, 50, 100 and 150 μM H₂O₂, respectively for 6 hrs. Caspase 3/7 activity was measured by using luminogenic substrate containing the DEVD sequence. Error bars indicate S.D., n=3, *P<0.05 vs. WT treated with H₂O₂.