cis-polymorphisms affect the level of precursor 80 kDa GALC. A, Lysates of cells transfected with the GALC mutants and cis-polymorphisms were analyzed by Western blot with anti-FLAG antibody, which detected only the 80 kDa precursor form. The FLAG antibody did not detect the 30 kDa fragment, suggesting that the FLAG tag at the C terminus might slightly modify the 30 kDa structure and mask the epitope. B, Quantification of the 80 kDa GALC densities showed that the c.550T polymorphism increased the unprocessed WT GALC, whereas none of the polymorphisms significantly affected the unprocessed G286D GALC. Both c.1685C and c.742A polymorphisms reduced the unprocessed level of T529M GALC. The polymorphism c.1685C did not have much effect on the unprocessed Y567S GALC. Error bars indicate SD of triplicates. *p < 0.05 (Student's t test). **p < 0.01 (Student's t test). ***p < 0.001 (Student's t test).