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Figure 4 Immunoblot of Kv3.1b stable expressed in GnT-I knockout NB cells. (A) A Western blot of glycosylated (WT) and unglycosylated (N220/229Q) Kv3.1b proteins in total membranes from stably transfected NB_1(-Mgat1) cells. (B) A Western blot of total membrane proteins from NB_1(-Mgat1) cells stably expressing WT Kv3.1b digested (+) and undigested (−) with PNGase F and Endo H. (C) A Western blot of Kv3.1b from NB_1 (−Mgat1) cells stably expressing WT Kv3.1b compared to those cells transiently transfected with GnT-I. Molecular weight standards in kDa: 250; 150; 100; 75; 50; and 37 from top to bottom. Black and grey arrows denote Kv3.1b glycoprotein with oligomannose and complex types of N-glycans, respectively. The grey line signifies unglycosylated Kv3.1 protein. See also Figures S6–S8.