Figure 2
PRDX6 upregulation is not strictly dependent on PrP expression. (A) Lysates of PrP-deficient (PrP0/0) and PrPC-expressing cells (PrP) were analyzed for PrPC, PRDX6 and β-actin. (B) Semi-quantitative PCR was performed with specific primers for murine prdx6, prnp and gapdh with (+) or without (−) reverse transcriptase (RT). (C) PrP0/0 and PrP cells were transfected with control siRNA (Ctr. siRNA) or siRNAs specific for murine PrP (PrP siRNA). 72 h after transfection cell lysates were prepared and Western blot analysis using the specific anti-PrP antibody SAF32 and anti-PRDX6 antibody were performed. β-actin was shown as a loading control. (D) Cells were incubated with 5 μg/ml cycloheximide and lysed after indicated time points. Equal amounts of cell lysates were separated by SDS-PAGE and detection of PRDX6, PrP and β-actin was carried out using specific antibodies