FIG. 6.

Degradation of FAT10 is ubiquitin independent. (A) The E1 thermosensitive cell line E36-ts20 (−E1) and E36-ts20 cells retransfected with a wild-type E1 (+E1) were transiently transfected with HA-tagged FAT10-AV-GFP. After inactivation of the thermolabile E1 by temperature shifting, the cells were labeled for 1 h with [³⁵S]Cys-Met and chased for the indicated time periods prior to HA-specific immunoprecipitation, SDS/PAGE, and autoradiography. (B) Similar experiments were performed transfecting a short-lived HA-DHFR-ubiquitin-Arg-β-Galactosidase construct. The immunoprecipitation was performed with an anti-β-Gal antibody. (C) The bands were quantified on a radioimager and plotted as percent radioactivity based on values obtained directly after the pulse. The means of results from three independent experiments are shown, and error bars indicate the standard errors of the means.