FIGURE 8.

Western blot analysis of PLN lysates with S2 and MECA-79. A, immunoblot of lysates (20 μg) from WT or GlcNAc6ST-1 and GlcNAc6ST-2 DKO (ST DKO) mice probed with 0.5 μg/ml biotinylated MECA-79 or -S2. B, immunoblot of lysates from WT or C1β3GnT and C2GnT-I DKO (C1/C2 DKO) mice treated with buffer alone (−) or with N-glycosidase F (+) probed with 0.5 μg/ml biotinylated E-PHA, -MECA-79, or -S2. N-glycosidase F treatment (100 units/ml; Roche Applied Science) was performed in PBS containing 0.9% Triton X-100 and a protease inhibitor mixture (1:40 dilution; Sigma-Aldrich) for 2 h at 37 °C. Ten micrograms of lysate proteins were applied to each lane, except that 100 μg of lysate proteins from the C1β3GnT and C2GnT-I DKO mice were applied for the blot with S2. The data are representative of three independent experiments.