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. 2017 Dec 13;9(4):4798–4813. doi: 10.18632/oncotarget.23219

Figure 5. PTCH1 and SLC39A6 were direct targets of miR-101-3p.

Figure 5

(A) Venn diagrams for miRNAs binding to both PTCH1 and SLC39A6 using two open-access, including miRDB and CLIP-seq database. (B-C) Sequence alignment of human miR-101-3p and 3’-UTR of PTCH1 and SLC39A6 using TargetScan and microRNA.org. The seed sequence of miR-101-3p (top) matches 3’-UTR of PTCH1(B, middle) and SLC39A6(C, middle). Bottom is the mutations of the 3’-UTR of PTCH1 and SLC39A6 used in luciferase reporter construct. (D-E and G-H) RT-PCR and western blotting analysis of PTCH1 and SLC39A6 mRNA and protein expression levels after overexpression miR-101-3p in A549 and H1299. GAPDH was used as internal control. (F, I) MiR-101-3p overexpression decrease the luciferase activity of PTCH1 and SLC39A6, but did not affect that of PTCH1-mut and SLC39A6-mut. Data are presented as the mean ± SD (n = 3). Significance was defined as p<0.05 (*, p < 0.05; **, p < 0.01; ***, p < 0.001).