Figure 4.

Serum deprivation increases the expression of Bim, a target of miR-101-3p. Cells were cultured in 5, 1, and 0% FBS-supplemented media in the presence of 50 μM Z-VAD-fmk for 16 h. (a and b) Bim mRNA and protein levels were determined at different time points by qRT-PCR and western blot analysis. (c and d) Cells transfected with C-miR, miRNA-101, antagomiR-101-3p or psiCHECK-2/Bim 3′-UTR (wild-type or mutant) were cultured in 5% FBS-supplemented or serum-free media containing 50 μM Z-VAD-fmk for 16 h. (c) Bim 3′-UTR activity was determined using a dual-luciferase reporter assay system. (d) Bim protein level was determined by western blot analysis. Bim_EL, Bim_L, and Bim_S indicate extra long, long, and short forms of Bim. **P<0.01