Figure 1.

Doxycycline induction of miRNA correlates with increased nuclear YFP and reduced luciferase reporter levels. (A) Schematic of constructs stably integrated in U2OS cells. Doxycycline induction produces intronic miRNA targeting the 3′-UTR of a luciferase reporter mRNA. Nuclear YFP is also expressed as an indirect readout of miRNA production. (B) The majority of cells produce significant levels of nuclear YFP by 12 h of doxycycline induction. Threshold is 3× mean background levels of YFP expression before doxycyline induction (0 h). N ∼ 300 cells, three replicates per data point. (C) Doxycycline induction results in reduced luciferase activity. N = 3 replicates per data point. Error bars = standard deviation.