FIGURE 2.

Detection of HNK-1-carrier proteins in NSCs and differentiated cells. a, lysates prepared from primary (lane 1), secondary (lane 3), and tertiary (lane 5) neurospheres and cells differentiated from primary (lane 2), secondary (lane 4) and tertiary (lane 6) neurospheres were analyzed by Western blotting with NGR50 anti-HNK-1 antibody or anti-β-actin antibody (Blot). β-Actin was detected as a loading control. Arrow indicates a major band positive for HNK-1 with a molecular mass of 280 kDa. b, lysates of primary neurospheres and their differentiated cells (containing 20 μg of proteins) were incubated with PNGase F (0 or 50 units/20 μl) at 37 °C for 3 h to remove N-glycans and then analyzed by Western blotting with anti-HNK-1 and anti-β-actin antibodies. undiffer and differ indicate lysates from primary neurospheres and differentiated cells, respectively.