Fig. 6

Competitive binding of insulin-like growth factor 2 messenger RNA binding protein (IMP1) to urethral carcinoma-associated 1 (UCA1) decreases the binding of miR-122-5p to UCA1. a Lentivirus vector expressing UCA1-MS2 was infected into MDA231/IMP1-green fluorescent protein (GFP) and MDA231/GFP cells. MS2 pulldown followed by RT-PCR analysis indicates the precipitated UCA1-MS2. b Enrichment of miR-122-5p in the precipitates of UCA1-MS2 was measured by RT-qPCR. IMP1 expression decreased binding of miR-122-5p to UCA1: **P < 0.01 as determined by Student’s t test. c MS2 pulldown analysis showed that UCA1 with muatated IMP1 binding motifs does not bind to IMP1. d RT-qPCR indicates that the UCA1-IMP1 interaction decreased the association of UCA1 with miR-122-5p. e Levels of pyruvate kinase muscle isozyme M2 (PKM2), insulin-like growth factor 1 receptor (IGF-1R) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) messenger RNAs (mRNAs) were measured in MDA231/UCA1, MDA231/UCA1-small interfering RNA (siRNA) and MDA231 control cells. Relative levels of mRNAs are the means ± SD from three independent experiments: **P < 0.01, *P < 0.05 as determined by one-way analysis of variance followed by Tukey’s multiple comparison tests. f RT-qPCR was used to monitor the effect of IMP1 overexpression on the expression levels of PKM2 and IGF-1R mRNAs. GAPDH mRNA was used as an internal control; means ± SD from three independent experiments: **P < 0.01 as determined by Student’s t test