Regulation of miR-146a by RelA/NFkB and p53 in STHdhQ111/HdhQ111 Cells, a Cell Model of Huntington's Disease
Figure 1
RelA/NFkB expression and activity in STHdhQ111/HdhQ111 and STHdhQ7/HdhQ7 cells.
(A) Representative Western Blot showing decreased level of RelA/NFkB (denoted by p65 in the figure) in STHdhQ111/HdhQ111 cells compared to STHdhQ7/HdhQ7 cells; (B) Average integrated optical density (IOD) of p65 protein bands (RelA/NFkB) in A, normalized to β-actin level (n = 3, p = 0.018) in these cells; (C) Average luciferase activity using reporter luciferase with multiple NFkB response elements (denoted by NFkB-RE) in STHdhQ111/HdhQ111 cells compared to STHdhQ7/HdhQ7 cells. Normalization of protein level between STHdhQ7/HdhQ7 cells and STHdhQ111/HdhQ111 cells was done by taking the ratio of Relative Luciferase Units (RLU) of NFkB-RE and empty vector pGL3 in these cells. The normalized value obtained with STHdhQ7/HdhQ7 cells was taken as 1. Relative luciferase activity of NFkB-RE was found significantly lower (n = 4, p = 0.0082) in STHdhQ111/HdhQ111 cells compared to STHdhQ7/HdhQ7 cells; (D) Average reporter luciferase activity with Gastrin promoter (n = 2, p = 0.034) in STHdhQ7/HdhQ7 cells and STHdhQ111/HdhQ111 cells. Normalization of protein level between the cells was done by taking the ratio of Relative Luciferase Units (RLU) of Gastrin promoter construct and empty vector pGL3 in these cells. The normalized value obtained with STHdhQ7/HdhQ7 cells was taken as 1; Error bars represent standard deviation s of more than 2 experiments and each experiment was done in duplicate. “*” represents statistical significance; * p≤0.05; ** p<0.01.