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. 2012 Dec 17;7(12):e51849. doi: 10.1371/journal.pone.0051849

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© 2012 Zheng et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Effects of siRNA-mediated Mpl knockdown. Dami cells were transfected with Ad-ShRNA-Mpl. Total cell proteins were prepared at the indicated times, and the effects of Ad-ShRNA-Mpl transfection on Mpl expression levels were evaluated by immunoblotting using an anti-Mpl antibody. Determination of the protein levels of β-Actin (bottom) was used as a control to ensure equal protein loading. (B) Time course of hNUDC released into culture medium after Dami cells were co-transfected with Ad-hNUDC and Ad-ShRNA-Mpl. (C) Comparison of hNUDC released into culture medium in cells transfected with Ad-hNUDC and control Ad-shRNA-scramble. (D) Time course of hNUDC released into culture medium after Dami cells were co-transfected with Ad-hNUDC(1-159) and Ad-ShRNA-Mpl. (E) Comparison of hNUDC(1-159) release into culture medium in Dami cells transfected with Ad-hNUDC(1-159) and control Ad-shRNA-scramble. Proteins released into the culture medium were analyzed by immunoblot using an anti-hNUDC antibody at each time point as indicated. Results are from 3–5 independent experiments.