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. 2017 Jan 31;8(26):42043–42060. doi: 10.18632/oncotarget.14927

Figure 6. The simultaneous expression regulation of target genes by miR-1.

Figure 6

A. The time-course assays of the miR-1-guided cleavage of target mRNAs. Each target mRNA of miR-1 was incubated with miR-1 and the Ago2 complex for various times at 37°C. The mRNA alone was used as a control. The cleavage products were examined using Northern blot analysis. The probes are shown at the top, and the numbers indicate the cleavage time. B. The effect of miR-1 concentration on target cleavage. Each miR-1 target mRNA was incubated with the Ago2 complex and different concentrations of miR-1 for 1 h. Then the cleavage products were detected by Northern blot analysis. The mRNA alone was used as a control. C. The miR-1-guided simultaneous cleavage of miR-1 target mRNAs. The mRNAs of all six miR-1 targets were equivalently co-incubated with the Ago2 complex and miR-1. One hour later, the cleavage products were examined by Northern blot analysis. The mRNA alone was used as a control. D. The intracellular co-localization of miR-1 and its six target genes. MDA-MB-435 cells were transfected with the miR-1 precursor and then cultured for 48 h. The cells were subjected to fluorescence in situ hybridization using a DIG-labeled miR-1 probe and a biotin-labeled mRNA probe. Confocal microscopy images are presented. Green, target gene mRNA. Red, miR-1. Blue, nuclei. Scale bar, 10 μm.