Figure 3

Figure 3

The effect of various point mutations in Eno2 on promoting the neurite growth of motor neurons. (A,B) Morphological differentiation of cultured NSC34 neural cells: (A) one-day incubation; (B) five-day incubation. Axonal neurites derived from cultured NSC34 cells were observed, and the neurite length was measured. (C) Statistical analysis of the average length of neurites. NSC34 cells transfected with siRNA (pCS2⁺) to inhibit endogenous mouse eno2 served as a negative control. After transfection with eno2 siRNA, eno2 DNA with modified wobble-nucleotides (eno2-wb) was transfected in NSC34. The resultant neurite length was normalized to a value of 1.0, which served as the positive control. The average neurite length of motor neurons was measured after transfection with siRNA, followed by transfection with plasmid harboring different point-mutated eno2-wb DNA. The fold change in neurite length was calculated relative to the positive control set as 1.0. Then, each experimental group was independently compared with the control group using t-test for statistical analysis (* p < 0.05, ** p < 0.01; ns indicates no significant difference). The t values and degrees of freedom for groups of -[L410I], -[M411L], -[D419S], -[E420K], -[R422K], -[H426R] and -[N427S] were 0.5754 and 2, 2.309 and 2, 6.362 and 3, 2.064 and 3, 5.413 and 2, 4.255 and 2, and 4.496 and 3, respectively.