Fig. 4.

SLC34A2 is the direct target of miR-939. a predicted miR-939 target sequence in 3'UTR of SLC34A2 (SLC34A2-3′UTR) and mutant containing three altered nucleotides in the seed sequence of miR-939 (miR-939-mut). b luciferase assay of pGL3-SLC34A2-3′-UTR reporters cotransfected with increasing amounts (10, 20, and 50 nM) of miR-939 mimic and mutant oligonucleotides in GC cell lines, or with increasing amounts (20, 50, and 100 nM) of miR-939 inhibitor oligonucleotides in GC cell line. c real-time RT-PCR analysis showed that overexpression of miR-939 significantly suppressed SLC34A2 mRNA expression level of SLC34A2 of GC cells, while inhibition of miR-939 led to a noticeable increase in SLC34A2 mRNA expression level of GC cell. d western blot analysis demonstrated that miR-939 tranfection decreased SLC34A2 protein level of GC cells, while anti-miR-939 dramatically increased SLC34A2 protein in GC cell. e high SLC34A2 IHC staining was observed in one GC sample with low miR-939 expression (upper panel); one GC sample with high miR-939 expression showed weak GNA13 IHC staining (lower panel). f the expression of SLC34A2 was inversely correlated with the expression of miR-939 in 112 GC samples. * P < 0.05, **P <0.001