Ube2A and UbcH5b localize to aggregates. The localization of Ubc's in cells containing polyglutamine aggregates was determined by immunofluorescence. HEK293 cells were cultured in 24 well plates, fixed and immunostained with antibodies against Ube2A, UbcH5b and E2-25K proteins. Control slides lacked primary antibody. Approximately 150 cells per Ubc enzyme were observed. To quantify colocalization, the percent of cells which contained YFP aggregates that also stained with the Ubc antibody was determined. In Ube2A and UbcH5b there was partial localization (49% and 71% respectively) whereas E2-25K showed no distinct colocalization with aggregates.
Howard et al. BMC Cell Biology 2007 8:32 doi:10.1186/1471-2121-8-32