Figure 5
Development of miR-9-5p and miR-9-3p sensors using destabilized CFP and Venus. (a) HeLa cells transfected with dsCFP, VP, or dsVenus were treated with 100 µg/mL CHX and analyzed by western blotting with anti-GFP and anti-β-actin antibodies. (b) Structure of miR-9-5p and miR-9-3p sensors. (c) Stable HeLa cells expressing both dsVenus-9-5p-T and dsCFP-9-3p-T were transfected with 1 nM miR-9-5p, miR-9-3p, or control miRNA mimics and were analyzed by western blotting with anti-GFP antibody. (d) Stable HeLa cells expressing both dsVenus-9-5p-T and dsCFP-9-3p-T were treated with 100 µg/mL CHX or 0.1% DMSO. Fluorescence intensities of dsVenus-9-5p-T and dsCFP-9-3p-T were analyzed by time-lapse imaging and are presented as mean ± SEM. Numbers of cells analyzed are shown in parentheses.
