Figure 6.

PRDX3 expression impacts sensitivity to oxidative stress and MMC independent of FANCD2 monoubiquitination. (A) FA-G mutant (PD352-F1°) and corrected (PD352-F2) fibroblasts were transfected with the pEF6/His6 vector (empty vector [EV]) or pEF6-PRDX3 (PRDX3). 24 h later, cells were exposed to H₂O₂ for 90 min, and their viability was assessed with an MTT assay. A representative experiment is shown with the line going through the mean and error bars showing the minimum and maximum value for each cell line. (B) HeLa cells were transfected with control siRNA (white bars) or PRDX3 siRNA (black bars). 24 h later, cells were exposed to MMC for an additional 24 h, and the number of viable cells was determined by MTT assay. (C) HeLa cells were transfected with control or PRDX3 siRNA and subsequently treated with 40 ng/ml MMC for 24 h. Protein lysates were made, electrophoresed, and immunoblotted with antibodies to PRDX3, FAND2, and KU70. The long (L) and short (S) FANCD2 species are noted.