Volume 7 issue 11 November 2006
ULTRASTRUCTURAL ANALYSIS OF ESCRT PROTEINS SUGGESTS A ROLE FOR ENDOSOME-ASSOCIATED TUBULAR-VESICULAR MEMBRANES IN ESCRT FUNCTION
Sonja Welsch, Anja Habermann, Stefanie Jäger, Barbara Müller, Jacomine Krijnse-Locker, Hans-Georg Kräusslich

Supplement 1:
Overexpression of Vps4_E228Q –EGFP leads to a redistribution of ESCRT in HeLa cells, but does not change overall labeling density of ESCRT proteins. HeLa cells were either transfected with a Vps4_E228Q –EGFP construct or left untransfected (control) and thawed cryosections were labeled with either anti-Tsg101 and anti-Hrs antibodies.
(A) The average labeling density per section profile with anti-ESCRT antibodies does not change upon overexpression of dnVps4-EGFP. Total numbers of gold particles (representing anti-Tsg101 or anti-Hrs labeling) per cell profile were determined. Values represent counts on at least 10 transfected and 10 control profiles per antibody. Error bars denote standard deviation.
(B) Quantification of the redistribution of ESCRT proteins in Hela cells upon Vps4 _E228Q –EGFP overexpression. Gold particles, representing anti-Tsg101 or anti-Hrs labeling were quantified on the plasma membrane, endosomes and tubular-vesicular membranes of transfected and control cells. At least 200 gold particles were counted per antibody. Values were collected in two different labeling experiments and two different grids per experiment.

Supplement 1 (.jpg)