Fig. 9. ABCG2-K86M is glycosylated to the same degree as ABCG2-wt. (A) Deglycosylation of ABCG2 protein with PNGase F or endonuclease H (Endo H). Total cell lysates of ABCG2-wt or ABCG2-K86M were treated with denaturation buffer for 10 minutes at 100°C. 15 µg total protein were incubated for 2 hours at 37°C with 100 mU enzyme. Samples were analyzed by SDS-PAGE and western blotting using the monoclonal anti-ABCG2 antibody BXP-21. (B) Positive control for endonuclease H activity. Immature IL2R (at 35-40 kDa) is sensitive to endonuclease H and is fully deglycosylated whereas the mature IL2R (50-60 kDa) is insensitive to endonuclease H treatment.