FIG 2.

Ubiquitination occurs primarily on complex glycosylated CD133. (A) HEK293T cells were transiently transfected with a Flag (control) or CD133-Flag plasmid. IP methods were performed to purify CD133 protein. PNGase F and endo H were applied for deglycosylation of CD133. PHA-L and ConA were used to examine complex glycosylated CD133 and high-mannose glycosylated CD133, respectively. (B) U87MG cells were used to stably express Flag or CD133-Flag. CD133 was precipitated using anti-Flag antibody. Complex glycosylated CD133 and high-mannose glycosylated CD133 were monitored by use of PHA-L and ConA, respectively. Red arrows indicate complex glycosylated CD133. Blue arrows indicate high-mannose glycosylated CD133. All results were collected from three independent experiments. Exp., exposure; IP, immunoprecipitation.