Fig. 4

MiR-335 activates WNT signaling through targeting APC and AXIN-I. a Pairing status of miR-335-3p and miR-335-5p with MREs within 3′UTR sequences of APC and AXIN-I. b Alterations in the expression of miR-335-3p did not significantly change the transcription level of APC gene (top); however, dual luciferase assay supported a direct interaction between them (bottom). c Neither RT-qPCR nor dual luciferase assay showed an interaction between miR-335-5p and APC. d Alterations in the expression of miR-335-3p significantly changed the AXIN-I transcript level (top), and dual luciferase assay indicated a direct interaction between them (bottom). e Although RT-qPCR suggested downregulation of AXIN-I following miR-335-5p overexpression (top), dual luciferase assay did not show an interaction between them (bottom). f Overall inductive effect of both miR-335-3p (top) and miR-335-5p (bottom) on WNT signaling as detected by TOP/FOP flash assay. All experiments were done in three biological replicates and results are presented as mean ± SEM