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. 2014 Oct 8;289(48):33296–33310. doi: 10.1074/jbc.M114.578245

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — STAM1 expression leads to a diffuse localization of Hrs and a recovery from the endosome enlargement induced by Hrs overexpression. A, HA-Hrs-transduced HEK293T cell line, 2HP67, containing enlarged early endosomes. 2HP67 cells were transfected with GFP-SARA-FYVE (GFP-FYVE) as an early endosome marker and then immunostained for HA-Hrs. B, endogenous ESCRT-0 proteins in HEK293T cells. C, endogenous STAM1 localizes to Hrs-positive microdomains within enlarged endosomes. 2HP67 cells were immunostained for HA-Hrs and endogenous STAM1. D, STAM1 expression in Hrs-overexpressing cells results in a diffuse localization of Hrs and the disappearance of enlarged endosomes. 2HP67 cells were transfected with FLAG-STAM1 and then immunostained for HA-Hrs and FLAG-STAM1. E, morphology of early endosomes under the same experimental conditions as that of D. 2HP67 cells were transfected with FLAG-STAM1 and then immunostained for EEA1, as an early endosome marker, and HA-Hrs. F, overexpression of STAM1 has no effect on the morphology of EEA1-positive endosomes. HEK293T cells were transfected with FLAG-STAM1 and then immunostained for EEA1 and FLAG-STAM1. Scale bars, 5 μm.