Fig. 4.

MicroRNA-27b and mmu-miR-298 negatively regulate VDR/NR1I1 expression. A, representative Western blot analyses indicated that LS-180 and PANC1 cells transfected with pS-miR-27b or pS-mmu-miR-298 plasmid had lower VDR protein expression. B, quantitative real-time PCR showed that lower VDR protein expression caused by miR-27b was associated with reduced CDS and 3′UTR mRNA levels. ∗, P < 0.05, compared with the corresponding control (n = 3 in each group, which refers to the number of independent transfection samples in a representative experiment; one-way ANOVA with Dunnett's posttest). Whole-cell proteins were separated by SDS-PAGE, and immunoblot analyses were carried out with selective antibody against VDR and GAPDH, respectively. See Materials and Methods for more details.