FIG 3.

Conversion of pre-miRNA to mature miRNA during induced expression following lentiviral transduction. (A) 293T and Neuro-2a cells were stably transduced with an inducible pre-miR-H2 expressing lentivirus, resulting in 293TH2 and Neuro-2aH2 cell lines. After 1 μg/ml of Dox was added, cells were harvested at different times and analyzed by Northern blot hybridization for miR-H2 expression. Cell line names and time points are indicated at the tops of the images. Positions of miRNA species are labeled to the left. Ratios of band intensities of pre-miRNAs to those of miRNAs are shown at the bottom. The experiment using 293TH2 cells was performed twice; the experiment using Neuro-2aH2 cells was performed once. (B) As described for panel A, but the 293TH3/H4 and Neuro-2aH3/H4 cell lines were created by transduction using a lentivirus expressing miR-H3 and miR-H4, and miR-H3 and H4 expression levels were analyzed. These experiments were repeated with cells constructed using different lentivirus constructs expressing miR-H3 and/or miR-H4.