Fig. 6.

OA induces pri-miR-7-1 processing in vitro. (a and b) In vitro processing of pri-miR-7-1 and (c and d) pri-miR-16 with HeLa extract in the presence of different concentrations of OA and EA. The processing of pri-miR-7-1 is increased upon (a) OA and (b) EA treatment. Radiolabeled pri-miR-7-1 (~ 30 × 10³ cpm) transcript was incubated with 50% (wt/vol) total HeLa cell extract, and the concentration-dependent effect of (a) OA and (b) EA treatment was assessed on the processing of pri-miR-7-1. The in vitro processing of pri-miR-16 is inhibited upon (c) OA and (d) EA treatment. Radiolabeled pri-miR-16 (~ 30 × 10³ cpm) was incubated with the 50% (wt/vol) total HeLa cell extract, and the concentration-dependent effect of (a) OA and (b) EA treatment was assessed on the processing of pri-miR-16. All the products of in vitro transcription were resolved on 8% denaturing polyacrylamide gel. Lane 1 in each gel represents the decade marker for RNA size. Lanes 2 and 3 in each gel represent mock treatment and in vitro processing reactions, respectively. Lanes 4 and 5 show in vitro processing reaction in increasing concentrations of OA and EA as indicated in the figure. These experiments were repeated for a minimum of three times.