Figure 5.

miR-146b Overexpression Promoted ETS2 Expression by Stabilizing Its mRNA

(A) T24T (miR-146b inhibitor) and T24T (nonsense) cells were cultured in 6-well plates until cell density reached 70%–80%. Following synchronization for 12 h, the medium was then replaced with 5% FBS DMEM-Ham’s F12 for another 12 h, and the cells were extracted for total RNA with Trizol reagent. Real-time PCR was used to determine ets2 mRNA expression, and β-Actin was used as an internal control. Data represent mean ± SD (*p < 0.05). (B) Human ets2 promoter-driven luciferase reporter was used to evaluate its promoter transcription activity in the indicated transfectants. The results were normalized by internal TK activity. (C) T24T (miR-146b inhibitor) and T24T (nonsense) cells were seeded into 6-well plates. After synchronization, the indicated cells were treated with Actinomycin D (Act D) for the indicated time points. Total RNA was then isolated and subjected to real-time PCR analysis for mRNA levels of ets2, and β-Actin was used as an internal control. The error bars represent mean ± SD from three independent experiments. Student’s t test was utilized to determine the p value. The asterisk (*) indicates a significant decrease in comparison to T24T (nonsense) cells (*p < 0.05).