Transfection of shRNA plasmids effectively suppresses PrP^C mRNA transcript and protein levels in murine N2a cells.

<p>(A) Schematic showing the relative locations of the target sequences in the 3′ untranslated region of the murine <i>prnp</i> locus. Black, dotted and grey arrows designate target sequences of p1578, p1633 and p1672, respectively. (B) Quantitative RT-PCR shows the relative mRNA expression in N2a cells transfected with the indicated amounts of p1578 (black bars), p1633 (checkered), p1672 (grey), pControl (stippled) and empty vector (striped) relative to untransfected control cells (white). (C) FACS analysis of cell-surface PrP^C expression in transfected cells. Peak fill colors correspond to the bar colors shown in (B). The light grey peak represents unstained control cells. (D) Cumulative FACS data analysis. Bar colors correspond to the same sample groups shown in (B). All three shRNA plasmids reduced PrP^C transcript (B) and protein (C and D) expression in N2a cells in a dose-dependent manner. Data from each assay were collected from at least three independent experiments.