(A) In the cells with neuroblastoma protective allele and higher NEFL expression levels, NEFL transient over-expression leads to inhibition of cellular growth. (B,C) In cells homozygous for the risk allele that have low NEFL expression, the growth inhibition is less marked. Detection of NEFL protein by using antibody against Flag and NEFL assessed the NEFL transiently overexpression in SH-SY5Y, SK-N-AS, IMR32 cell lines. The variability of the forced NEFL expression may to be due to the different transfection efficiency of the three different cell lines. (D,E) We thus produced a stable over-expression of NEFL in SH-SY5Y and SK-N-AS cells to obtain a further validation. The experiments have been performed in three diverse clones for each experimental point and the shown results are representative of the mean among the different clones. The numbers represent the cell number difference between control and NEFL over-expression. Times (T) represent the hours.