Western blotting results of Elk1 and Elk1-3R effects on both endogenous katanin-p60 and spastin proteins.

(A) Analysis of SH-SY5Y transfection efficacy with His-tag including pCMV6_Elk1 and pCMV6_Elk1-3R vectors. (B) Western blotting results showing katanin-p60 level in untransfected (control), pCMV6_Elk1 transfected, and pCMV6_Elk1-3R transfected SH-SY5Y cells. Transfection experiment was performed two times on separate days and Western blotting experiment was performed two times with each sample. (C) Quantification of katanin-p60 level was performed by normalizing band intensities of katanin-p60 to band intensities of ß-actin. Results indicated that both Elk1 and Elk1-3R decreased the katanin-p60 level significantly and the difference between them was not significant. SEM values are 0, 0.01, and 0.02, respectively (n = 4). (D) Western blotting results showing spastin level in untransfected (control), pCMV6_Elk1 transfected, and pCMV6_Elk1-3R transfected SH-SY5Y cells. Transfection experiment was performed two times on separate days and Western blotting experiment was performed two times with each sample. (E) Quantification of spastin level was done as previously described. Results indicated that there was no significant spastin level difference between Elk1 and Elk1-3R transfected cells. However, they were significant compared to untransfected cells. SEM values are 0, 0.03, and 0.03, respectively (n = 4).