Katanin-p80 Gene Promoter Characterization and Regulation via Elk1
Figure 5
Elk1 activates KATNB1 promoter and increases katanin-p80 mRNA.
(A) Forced luciferase experiment to compare effects of Elk1 on KATNB1 optimal promoter activation. SH-SY5Y cells were co-transfected with (I) F1 construct and either pCMV-empty vector (F1) or pCMV6-Elk1 (F1+Elk1) and (II) F2 construct and either control (F2) or Elk1 (F2+Elk1). (III) Normalization of data is shown in I and II. Promoter activity of F1 and F2 constructs transfected with Elk1 containing vector were normalized to F1 and F2 constructs transfected with control vector, respectively. (B) Western blot image represents the total protein lysate obtained either from Elk1 transfected (overexpression) or naive SH-SY5Y cells (control). Cells were immunolabeled for Elk 1 and β-actin. (C) Relative mRNA quantification of katanin-p80 mRNA level in Elk1 overexpressing SH-SY5Y cells (Elk1). Results were calculated by normalizing the Ct values of the KATNB1 gene to β-actin as internal control. Results represent ΔΔCt values which gives mRNA fold change relative to naive cells (control). Error bars represent ± SD. Asterisk symbol (*) indicates p-value <0.05, meaning a statistically meaningful difference.