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. 2012 Feb 6;109(11):4146–4151. doi: 10.1073/pnas.1200448109

Fig. 3.

Fig. 3.

The ARRDC1–TSG101 interaction and the budding-related protein VPS4a are required for release of ARMMs. (A) ARRDC1 and TSG101 are required for ARMMs release. Western blot analysis of ARMMs and whole-cell extracts from HEK293T cells subjected to transfection by indicated siRNAs. The results shown are representative of three independent replication experiments. (B) PSAP-mutant ARRDC1 is released less efficiently into ARMMs. Cells were transfected with wild-type or PSAP-mutant (PAAP) ARRDC1-GFP, and the corresponding whole-cell extracts and ARMMs were analyzed by Western blotting probed by the indicated antibodies. (C) ARMMs release requires functional VPS4 ATPase activity. HEK293T cells were cotransfected with constructs expressing HA-ARRDC1 and 0.5 or 1 μg of VPS4a (VPS4) or VPS4a E228Q (VPS4-DN) GFP fusion proteins. Whole-cell extracts and ARMMs were analyzed by Western blotting using the indicated antibodies.