Figure 3.

Rapamycin inhibits miR-1 enhancer activity. (A) C2C12 cells were transfected with the miR-1-2 enhancer reporter or the corresponding enhancerless reporter, induced to differentiate (Diff) in the absence or presence of 50 nM rapamycin (Rap), and lysed for luciferase assays at the times indicated. (B) Cells were transfected as in A and induced to differentiate for 72 h in the presence of 50 nM rapamycin, 100 nM wortmannin (Wort), or 1 µM SB203580 (SB) followed by cell lysis and luciferase assays. The enhancerless reporter activity was subtracted from the enhancer reporter activity for each condition, and the data shown have been normalized with 0 h of activity as 1. Data shown are the mean ± SD of three independent experiments.