Fig. 6. Comparison of pro-NPP2 and NPP2. (A) HA-NPP2-Myc was expressed in HEK293 cells with or without the furin-inhibitor ₁-PDX, to generate pro-NPP2 and NPP2, respectively. After 24 hours, 48 hours and 72 hours the cell lysates and culture media were processed for immunoblotting with anti-Myc antibodies. (B) Pro-NPP2 and NPP2, obtained after the expression of HA-NPP2-Myc with or without the furin-inhibitor ₁-PDX, respectively, were purified on an anti-Myc affinity column. The purified enzymes were assayed for lysophospholipase-D activities. The bar diagrams represent the means ±s.e.m. (n=3) of the enzymatic activities. Pro-NPP2 and NPP2 were assayed at the same concentration, as measured in triplicate by immunoblotting with anti-c-Myc antibodies (inset).