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. 2020 Feb 13;20:97–110. doi: 10.1016/j.omtn.2020.02.006

Figure 2.

Figure 2

RC3H2 Directly Bound to miR-101-3p and Its Mainly Subcellular Localized to Cytoplasm

(A) qRT-PCR was used to determine the expression of miR-101-3p in HN4 and Cal27 cells transfected with si-RC3H2. (B) qRT-PCR was used to determine the expression of miR-101-3p in HN4 and Cal27 cells infected with LV-RC3H2. (C) qRT-PCR analysis of miR-101-3p following RNA pull-down assays with RC3H2 probes in HN4 and Cal27 cells. (D) StarBase version v2.0 results showing the sequence of RC3H2 with highly conserved putative miR-101-3p binding sites and construction of the RC3H2-Mut vector, which changed TACTGTAA into GCACACGA from nucleotides 518 to 525. miR-101-3p mimic considerably reduced the luciferase activity of the RC3H2-WT luciferase reporter vector compared with negative control, while miR-101-3p mimic did not have any impact on the luciferase activity of RC3H2-Mut-transfected cells. (E) FISH analysis of RC3H2 in HN4 cells. (The nuclei were stained with DAPI, and 18S rRNA was used as a cytoplasmic marker. Scale bars, 10 μm.) (F) FISH analysis of RC3H2 in Cal27 cells. (The nuclei were stained with DAPI, and 18S rRNA was used as a cytoplasmic marker. Scale bars, 10 μm.)