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. Author manuscript; available in PMC: 2019 Apr 3.
Published in final edited form as: Sci Signal. 2018 Apr 3;11(524):eaao4180. doi: 10.1126/scisignal.aao4180

Fig. 2. ORL1 enhances O-linked glycosylation of NgR1 at Golgi body.

Fig. 2.

(A) HEK293T cells were transfected with Myc-NgR1 or Myc-NgR2, with or without HA-ORL1. HA immunoprecipitates were immunoblotted for HA and Myc. Means ± SE, n = 3 independent experiments. (B) HEK293T cells were transfected with FLAG-NgR1 and increasing amounts of HA-ORL1 and treated without or with N-glycosidase or O-glycosidase for 1 hour. Cell lysates were immuno-blotted for FLAG or actin. Means ± SE, n = 3 independent experiments. glyco, glycosidase. (C) HEK293T cells were transfected with FLAG-NgR1, with or without HA-ORL1 and treated with dimethyl sulfoxide or increasing amounts of benzyl-N-acetyl-α-D-galactosaminide (b-GalNAc) as indicated. Cell lysates were immunoblotted for FLAG, HA, and actin. Means ± SE, n = 4 independent experiments. (D) Cos7 cells cotransfected with FLAG-NgR1 and HA-ORL1 were stained with NgR1, HA, and GM130 antibodies. Means ± SE, n = 3 independent experiments. Scale bar, 20 μm.