Figure 3

Silencing of b PRNP by siRNA as revealed by Western blot analysis. Top, protein size markers (Bio-Rad, lane 1), proteins from HeLaS3 cells (lanes 2–4), proteins from cells co-transfected with pbPrP- FLAG DNA and the control vector DNA of piGENE hU6 (lanes 5–7) or siRNA expression vector DNA of piGENE hU6-616-21, the sequence of which is No. 8 in Table 2 (lanes 8–10), proteins from cells co-transfected with pbPrP- FLAG DNA and the control vector DNA of piGENE tRNA (lanes 11–13) or siRNA expression vector DNA of piGENE tRNA-616-21(lanes 14–16), and protein size markers (150, 100, 75, 35, 25, and 15 kDa from the top, GE Healthcare). Amounts of proteins applied were 1.25 (lanes 2, 5, 8, 11, 14), 2.5 (lanes 3, 6, 9, 12, 15), and 5.0 (lanes 4, 7, 10, 13, 16) μg/lane. The gel was stained with SAF32 antibody. Bottom, actin staining.