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Figure S2. Endocytosis and dynamin protein levels are not affected in eps15-null hemocytes. (A-C) Maleylated BSA (mBSA) is endocytosed by both control and eps15^e75/eps15^Δ29 hemocytes. As described previously (Guha, A. V. Sriram, K.S. Krishnan, and S. Mayor. 2003. J. Cell Sci. 116:3373-3386), hemocytes harvested from third instar larvae were incubated with Texas red-conjugated mBSA for 1 min and in the absence of mBSA for a 4 more minutes. The assay was performed at 35^°C with control (Oregon R; A) and shi^ts1 hemocytes (C), and at room temperature for eps15^e75/eps15^Δ29 hemocytes (B). (D and E) Dynamin protein levels is similar between control (D) and eps15^e75/eps15^Δ29 hemocytes (E), as revealed by immunohistochemistry performed using rabbit anti-dynamin antibodies (Roos, J., and R.B. Kelly. 1998. J. Biol. Chem. 273:19108-19119). (F) Quantification of mBSA-TxRed uptake as determined in two independent experiments. All comparisons are with control; n = 3-7 larvae (7-17 cells) for each genotype. (G) Quantification of dynamin protein levels averaged per cell as determined in two immunofluorescence experiments; n = 6 larvae (22-29 cells) for each genotype. (F and G) Fluorescence intensity was quantified as mean luorescence per cell using ImageJ. The quantified data were processed and analyzed in Excel (Microsoft) using t test. No significant difference was found between control and eps15^e75/eps15^Δ29 hemocytes in F and G (P > 0.05).