Txnrd2

Western blot analysis was performed according to standard methods. Briefly, proteins were separated using 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then transferred to 0.45 µm polyvinylidene difluoride membranes (Merck Millipore, Germany), which were blocked in 5% nonfat milk for 1 h and incubated with primary antibodies at 4 °C overnight. After washing in a solution of tris-buffered saline and Tween 20, membranes were incubated for 1 h in corresponding secondary antibodies. Detection of the protein was conducted using a chemiluminescence kit (#P90719, Merck Millipore). β-Actin served as the loading control. The primary antibodies included: GSTP1 (#3369, Cell Signaling Technology, USA) and TXNRD2 (#ab181864, Abcam, UK). The dilution of primary antibodies was 1:1000. Secondary antibodies were goat anti-mouse or goat anti-rabbit IgG (H+L) poly-horseradish peroxidase secondary antibody (Jackson Immunoresearch, USA) and the dilution of the secondary antibodies was 1:10,000. Protein extracts from lens capsules of patients, cultured LECs, and whole lenses from experimental mice were used in this assay.