Hypoglycosylation of α-DG led to dramatically reduced laminin binding.
Glycoproteins were isolated from the whole brains and cultured neural stem cells by WGA-agarose, separated on SDS-PAGE, and transferred onto PVDF membranes. Pikachurin overlay and Western blot then were carried out. (A) Recombinant pikachurin (detected with anti-c-Myc) was detected in conditioned medium from HEK293 cells transfected with pikachurin expression plasmid. (B) Whole brain lysates. (C) Neural stem cells. Pikachurin binding was readily detected in the wildtype but dramatically reduced or abolished in Dag1f/f;Nestin-Cre(+), POMGnT1 knockout, and Largemyd mice and in POMT2-deficient neural stem cells. Abbreviations: β-DG, β-dystroglycan; KO, knockout.