Figure 9.
Functional cross-talk between p53 and NFκB p65/RelA regulate miR-100, −146a and −150 expressions in mouse striatal STHdhQ7/HdhQ7 cells Changes in relative luciferase activity of (A) miR-100 upstream sequence construct (B) miR-146a upstream sequence construct and (C) miR-150 upstream sequence construct in presence of exogenous p53-CFP and NFκB p65/RelA in mouse striatal STHdhQ7/HdhQ7 cells. In presence of exogenous NFκB p65/RelA luciferase activities of the constructs were increased whereas in presence of exogenous p53-CFP the luciferase activities of the constructs were significantly reduced; data are mean ± SD (n = 3); *P ≤ 0.05; ** P ≤ 0.01 compared to control. (D) Western blot showing increase in p65 protein level by more than 3 fold (*p≤ 0.05) and increase in p53 protein level by more than two fold († p≤ 0.05) when 2.5 μg each of p65 and p53 plasmid clones was transfected either in isolation or in combination in STHdhQ7/HdhQ7 cells compared to control STHdhQ7/HdhQ7 cells; (n = 3). (E) Chromatin immunoprecipitation assay showing decreased NFκB p65/RelA occupancy at its binding site in miR-100 upstream sequence (Reg C) in p53-CFP transfected STHdhQ7/HdhQ7 cells compared to control STHdhQ7/HdhQ7 cells, data are mean ± SD (n = 3); *P ≤ 0.05; ** P ≤ 0.01 compared to control. This indicates p53 driven exclusion of NFκB p65/RelA from its binding site that partially contributes to miR-100 repression. Error bars represent standard deviation, * represents statistical significance.