Figure 5

Prdx6 Knockdown experiments showing that Prdx6 deficiency contributed in the process of TM cell pathogenesis. (a) Normal TM cells were transfected with antisense-specific Prdx6 and enriched by selection with antibiotic. Cellular extracts from two groups (39Y and 54Y) of cells having equal amount of proteins were immunoblotted with anti-Prdx6, anti-TGFβ and ECM protein, anti-α-sm-actin antibodies by stripping and restripping the same membrane. Under expression of Prdx6 in these cells enhanced the levels of TGFβ, as well as α-sm-actin. (b–d) Prdx6 knockdown TM cells bore increased ROS levels with increased LPO contents and SA-β-Gal activity. TM cells under expressing Prdx6 were divided into three groups to assess (b), levels of ROS (c), contents of LPO levels and (d) levels of SA-β-gal activity, and were compared with empty vector transfected cells. Under expression of Prdx6 significantly increased ROS production (b), LPO (c) and SA-β-gal activity (d), suggesting that Prdx6 was indeed a cause of abnormalities in TM, and its deficiency increased pathobiological processes. Histogram values represent the mean±S.D. from three independent experiments (*P<0.001).