Fig. 1.

Scheme of the cell membrane N-glycomic workflow used for the characterization of region-specific cell surface mouse brain N-glycomes. After extraction, brains were surgically dissected either into forebrain and hindbrain (A) or the functional substructures derived from both regions, including the cortex and hippocampus (from forebrain) and cerebellum (from hindbrain) (B). Tissues dissected were homogenized, cell debris, nucleus, and mitochondria were removed by centrifugation from region-specific brain lysates. Cell membrane fractions enriched in cell surface glycoproteins were obtained by ultracentrifugation as previously described (38). Cell membrane glycans were enzymatically released, purified, and analyzed by nano-LC-Chip-Q-TOF mass spectrometry.