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. 2004 Aug 30;166(5):645–651. doi: 10.1083/jcb.200312072

Figure 4.

Figure 4.

Cytosolic delivery of DT is COPI dependent but does not require transport to late endosomes. (A and B) ldlF cells were grown at the permissive or restrictive temperature for 16 h, incubated for 7 h (A) or 2.5 h (B) at 37°C in serum-free medium with 500 ng/ml of DT or trypsin-nicked DT (DTn) or left untreated (cont.). (B) Cell lysates were prepared and the modification of EF2 analyzed as in Fig. 3 D. (C) Wild-type CHO cells were incubated or not with 10 μM nocodazole for 2 h at 37°C and 15 min at 4°C, followed by 1 h at 4°C with 500 ng/ml DTn, washed and further incubated for different times at 37°C. The modification of EF2 was analyzed as above. (D) CHO cells were treated with 6c4 antibody as in Fig. 3 A and incubated with 500 ng/ml DTn and modification of EF2 analyzed. The band labeled with an asterisk is a cross-reacting protein, apparent in certain experiments. (E) ALIX was knocked down in HeLa cells as in Fig. 3 B, incubated with 500 ng/ml DTn, and modification of EF2 analyzed.