FIGURE 3.

Prophenoloxidase activation cascade in the molting fluid. A, PO activities in larval-larval (LL), larval-pupal (LP), and pupal-adult (PA) molting fluids. Equal amounts of each molting fluid (10 μg) were used for PO activity assays. B–D, detection of PPO and βGRP-3 in molting fluids and integuments. Approximately 30 μg of cell lysates from integuments, various types of molting fluids, or 0.5 μl of plasma (V-2) was loaded per lane. In B–D, the arrow indicates the position of PPO (B and C) or βGRP-3 (D). Samples were the same as in supplemental Fig. S2. In C, molting fluids were kept at room temperature for 30 min before Western blot. E, H₂O₂ detected in molting fluids. Bars represent the mean of three independent measurements ± S.E. (error bars).