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. 2018 Nov 14;38(6):BSR20180982. doi: 10.1042/BSR20180982

Figure 4. RB1 is a direct target of miR-199a-3p.

Figure 4

(A) TargetScan analysis showing the WT 3′-UTR of RB1 mRNA containing a putative miR-199a-3p target site. A mutant (MUT) sequence was designed accordingly to be tested for luciferase assay together with the WT. (B) Luciferase reporter assay comparing WT with MUT RB1 3′-UTR targetting by miR-199a-3p. RA-FLSs cells were co-transfected with RB1 3′-UTR firefly luciferase reporter constructs harboring WT or MUT miR-199a-3p-targetting sequences and an miR-199a-3p-expressing plasmid (miR-199a-3p) or a pSliencer 4.1-CMV puro vector (miR-control). Firefly luciferase activity was normalized to Renilla luciferase activity. (C) RT-qPCR analysis of RB1 mRNA in RA-FLSs transfected with miR-199a-3p or miR-control. (D) RB1 protein expression in RA-FLSs transfected with miR-199a-3p or miR-control. n=5; *P<0.05 compared with miR-control.