Effect of over-expression of miR-22 in luciferase-expressing HEK cells.
A) Fold change (FC) of overall luciferase (blue), cell viability (orange), and luciferase per cell (grey) of Luc-HEK, Luc-HEK-miR-22, Luc-HEK-miR-22 pool and Luc-HEK-miR-NC cells relative to Luc-HEK demonstrates improved luciferase activity with microRNA 22 over-expression. Error bars represent Standard Error of the Mean (SEM) from triplicate measurements. * indicates P ≤ 0.05 relative to Luc-HEK and Δ indicates P ≤ 0.05 relative to Luc-HEK-miR-NC calculated using two-sample unpaired t-test assuming unequal variances. B) Western blot analysis confirms that over-expression of miR-22 improves luciferase expression in Luc-HEK-miR-22 (2.7 FC) and the Luc-HEK-miR-22 pool (2.9 FC) Luc-HEK-HIPK1 KO (2.4 FC), and the Luc-HEK-HIPK1 KO pool (1.5 FC) compared to Luc-HEK. A monoclonal antibody against firefly luciferase was used to detect the protein, with antibodies against β-actin as an endogenous control.