QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 3. N-glycosylation protects P2X₄ receptors from degradation in lysosomal compartments. (A) NRK cells transfected with vectors encoding either P2X₄ or GFP-tagged M4 muscarinic receptors were incubated with 5 µg/ml of CHX for the indicated durations. Thp-1 cells were similarly treated with CHX. Cell lysates were then analysed by SDS-PAGE and immunoblotted with antibodies against either GFP or P2X₄. (B) Lysates from NRK cells transfected with vector encoding P2X₄ were grown in the presence or absence of DMJ to prevent complex glycosylation and were treated with either 500 U Endo H or 12 U N-glycosidase F for 5 hours. (C) A DMJ-treated cell lysate was incubated with 125 U Endo H for 30 minutes to reveal six different glycosylation states. (D) NRK cells treated for 24 hours with DMJ (in the presence of 5 µg/ml of CHX) and Thp-1 cells treated for 3 days with DMJ were incubated with 50 mU/ml Endo H for the indicated number of hours. Cell lysates were then analyzed by immunoblotting with an antibody against P2X₄. (E) A similar experiment was performed with DMJ-treated RAW264.7 cells, and the lysates were analyzed by immunoblotting with antibodies against P2X₄ and LAMP-1.

Return to article