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Fig. 2. Efficiency of protein knockdown. (A) Representative images of cells immunostained with antibodies against endocytic proteins 3-4 days after RNAi (RNAi, bottom row) or cells left untreated (Control, top row). Maximum-intensity projections of several Z-sections, collected using fluorescent microscopy, are shown. (B) Protein levels in S2 cells after RNAi knockdown. Images similar to those in A were used to quantify average fluorescence per cell, data represent mean ± s.e.m. of three independent experiments. (C) Primary haemocytes (inside dotted circles) from eps15 and endoA mutant animals internalise mBSA normally. Haemocytes harvested from wild-type (WT), eps15 (EP(2)2513), endoA (endoA¹) and dynamin (shi^ts1) mutant third instar larvae (Guha et al., 2003) were incubated with mBSA-Texas-Red for 2 minutes, followed by a 4-minute chase. Assays were performed at 35°C with WT and shi^ts1 cells, and at room temperature with eps15 and endoA mutant cells.

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