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To specifically reduce RPTP mRNA abundance in these cells, we used short interfering RNA technology (siRNA) (Elbashir et al., 2001; Paddison et al., 2002). At 24 h following siRNA transfection, Western blot analysis of whole cell protein lysates using a RPTP specific antibody demonstrated a 73% reduction of RPTP protein. Reduction of RPTP expression was specific, as the unrelated sequence control siRNA had no detectable effect on RPTP expression compared to mock transfected cells (Figure 4a).
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