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Fig. 4. The N-terminus of NPP2 functions as a signal peptide. (A) The N-terminal sequence of NPP2 with the predicted N-terminal (n), hydrophobic (h) and C-terminal (c) fragment of the signal peptide sequence, as predicted by the Signal P program (version 3.1). Also indicated is the predicted cleavage site (arrow). (B) HA-NPP2-Myc, with a Flag epitope inserted after either Gly27 or Phe28, was expressed in HEK 293 cells. 24 hours after transfection the cells were harvested and the lysates were subjected to immunoblotting with anti-Flag M1 and M2 monoclonal antibodies. The M1 antibodies only recognize an N-terminally free Flag-tag, whereas the M2s also recognize an internal Flag-tag. The data are representative for at least three independent experiments.





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