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	Figure 2. Effect of different Rab proteins on HRP uptake in oocytes. Oocytes were injected with 100 ng of the indicated Rabs in 50 nl of microinjection buffer and incubated for 2 h at 18°C. The cells were then incubated with HRP (2 mg/ml) for 1 h at 18°C to determine the uptake rate. Results were expressed as percentage of uptake per oocyte with respect to control (220 ng per oocyte) ± SD (average of eight experiments).
	Figure 3. Effect of Rab5 and its mutants on endocytosis in oocytes. (a) Oocytes were injected with 100 ng each of the indicated proteins, and the uptake was carried out as described in Fig. 1. The results were expressed as percentage of control (200 ng/oocyte) ± SD. (b) Oocytes were injected with 100 ng of Rab5:Q79L or Rab5:ΔC4 in 50 nl of microinjection buffer and incubated for 2 h at 18°C. The cells were lysed, and crude membrane and cytosolic fractions were prepared by centrifuging at 70,000 rpm for 15 min. Each fraction (5 μg) was separated by SDS-PAGE and transferred onto nitrocellulose, and Rab5 protein was localized using polyclonal Rab5-specific antibodies (Alvarez-Dominguez et al., 1996).
	Figure 4. Inhibition of HRP uptake by anti-Rab5 antibody. The oocytes were injected with Rab5:Q79L (100 ng) alone or with anti-Rab5 antibody (200 ng). HRP uptake was followed as described in Fig. 1. Results are expressed as percentage of control uptake (150 ng per oocyte) from the average of eight experiments.
	Figure 5. Effect of NSF on HRP uptake by oocytes. (a) Cells were injected with either 100 ng of Rab5:Q79L with or without anti-NSF (200 ng) antibody. Uptake was carried out as described in Materials and Methods. Results were expressed as percentage of control uptake (180 ng/oocyte) from the average of eight experiments. (b) Oocytes were injected with 75 ng of Rab5:Q79L alone or with 100 ng of NSF and its mutants as indicated. Uptake was determined as described, and the results are expressed as percentage of control (220 ng per oocyte) uptake.
	Figure 6. Rab7-mediated uptake of HRP by oocytes. Oocytes were injected with 100 ng of the indicated proteins and uptake was carried out with HRP-FITC for 1 h in the dark (1 mg/ml). Subsequently, the cells were washed three times with MBS, incubated in the dark for another 1 h, and then observed under the confocal microscope.
	Figure 7. Rab7-mediated inactivation/degradation of HRP by oocytes. (a) Oocytes were preloaded with HRP by incubating with HRP (2 mg/ml) for 1 h at 18°C. Preloaded cells were washed and incubated with 100 μM monensin or control buffer as indicated for 1 h at 18°C. The cells were then injected with 100 ng of the respective fusion proteins. HRP activity present in the cells was measured after a 2-h incubation at 18°C. Results are expressed as percentage of control (110 ng HRP/oocyte) from the average of eight experiments. (b) Oocytes were incubated with 125I-HRP (0.5 mg/ml; 2 × 105 cpm/μg) for 1 h at 18°C and washed three times to remove unbound radioactivity. Subsequently, the cells were injected with 100 ng of Rab7:WT protein or control buffer, and nine oocytes were incubated in 300 μl of MBS for 2 h at 18°C. Oocytes were washed and solublized in PBS containing 0.1% Triton X-100, and TCA-soluble and -precipitable radioactivity was determined in cell lysates and media. Results are expressed as ng of HRP associated with or degraded by the cells.
	Figure 8. Rab5 regulation of endocytosis is upstream of Rab7. (a) Rab5 proteins were injected alone (50 ng) or in combination with both mutants (100 ng) of Rab5 and Rab7. (b) Rab7 proteins (50 ng) were injected alone or in combination with both mutants (100 ng) of Rab5 and Rab7. Results are expressed as percentage of control uptake by uninjected oocytes (average of eight experiments).
	Figure 9. Rab5- and Rab7-coinjected cells produced additive effects. Oocytes were injected with 50 ng of Rab5 and Rab7 alone or in combination and incubated for 2 h at 18°C. The cells were incubated with 2 mg/ml of HRP for 1 h at 18°C to determine the rate of HRP uptake. Results are expressed as percentage of uptake per oocyte with respect to control (205 ng per oocyte) ±SD (average of eight experiments).

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