FIG 7.

miR-199a-5p controls M6PR expression. (A) Luciferase reporter activity in COS7 cells transfected with control mimic (CM) or miR-199a-5p mimic and M6PR 3′ UTR containing or not containing (WT, wild-type) the indicated point mutations (PM) in the miR-199a-5p-binding sites. Data are expressed as percentages of M6PR 3′-UTR activities in miR-199a-5p- versus CM-transfected cells. *, P ≤ 0.05. DM, double mutation. (B) Quantitative real-time PCR analysis of M6PR expression in HeLa cells transfected with control mimic (CM) and miR-199a-5p mimic after 48 h. (C) Representative Western blot analysis of M6PR expression in HeLa cells treated as described in the legend to panel B. (B and C) Data are expressed as mean values ± SEM and are representative of the results of ≥3 experiments in triplicate. *, P ≤ 0.05. (D) Immunofluorescence analysis of Cy3-StxB internalization in HeLa cells cotransfected with CM and M6PR-GFP or miR-199a-5p and M6PR-GFP for 48 h. Steady-state localization is shown in Z projections of confocal stacks. Scale bar = 20 μm.