Figure 9

PRDX6 regulation of NF-kB expression and cellular survival during hypoxia (A). MTS assay showing protective effect of over-expression of PRDX6 on RGCs survival. Cells were transiently transfected with either GFP vector or GFP-PRDX6. After 24 h cells were either treated with CoCl₂ (50, 100 or 200 μM) or exposed to 1% O_2. Results indicate that GFP-PRDX6 over-expression significantly reduced death of RGCs exposed to CoCl₂ (A; 50, 100 or 200 μM) or 1% O₂ (B). To monitor the activation of NF-κB, cells were transfected with GFP vector or GFP-PRDX6. These cells were co-transfected with HIV-1LTR-CAT construct (C) having NF-κB sites [37] and were exposed to 1% O₂. Results showed the repression of NF-κB-dependent transcriptional activation of HIV-1LTR in cells over-expressing PRDX6 (C; black bar). In parallel experiments, cell extracts was prepared and Western analysis performed to confirm whether GFP-PRDX6 over-expression reduced the expression of NF-κB (D). Upper panel showing decreased expression of NF-κB in nuclear extract of RGCs over-expressing PRDX6.