Luciferase assay results of KATNA1 and SPG4.

(A) In order to ascertain the effect of Elk1 on KATNA1 regulatory regions, SH-SY5Y cells were co-transfected with pGL3-KPU construct and with either pCMV6_Elk1 or pCMV6_Elk1-3R. Also, pGL3-basic and pCMV6-empty vector co-transfection was performed. Luciferase assay was performed 48 h post-transfection, and calculated fold activities indicated that wt-Elk1 significantly decreased promoter + 5’ UTR activity of KATNA1. Even though Elk1-3R caused a partial increase compared to wt-Elk1, it was not statistically meaningful. Thus, it also significantly decreased promoter + 5’UTR activity compared to unforced (in the absence of Elk1) experiment. SEM values are 23.79, 10.99 and 11.92, respectively. Each experiment was performed as triplicates on the same day and the experiments were repeated three times on separate days, independently (n = 3). (B) For the same purpose, SH-SY5Y cells were transfected with pGL3-SP and with either pCMV6_Elk1 or pCMV6_Elk1-3R and pGL3-SP with pCMV6-empty vector. Both WT-Elk1 and Elk1-3R significantly decreased the promoter activity of SPG4. Even though Elk1-3R decreased the activity more than wt-Elk1, the difference was not significant. SEM values are 12.51, 8.41 and 9.83, respectively. Each experiment was performed as triplicates on the same day and the experiments were repeated three times on separate days, independently (n = 3).