Fig. 3

WWP1 was up-regulated in GC tissues and cells. WWP1 was proved to be the potential target gene of miR-584-5p. a. The expression level of WWP1 was detected in 75 pairs of human GC tissues and non-GC tissues by qRT-PCR. b. WWP1 protein level was determined using Western blot in six paired of GC tissues. GAPDH was used as the internal reference. c. The expression level of WWP1 protein in GC tissues and adjacent normal tissues was determined by immunohistochemistry staining. d. Negative correlation between the expression levels of miR-584-5p and WWP1 in GC specimens. e, f. The expression levels of WWP1 in GC cells and GES-1 were detected using qRT-PCR and Western blot. g. Luciferase reporter assay was conducted to verify that miR-584-5p directly bound to the 3’-UTR region of WWP1. Luciferase activity was analyzed in cells co-transfcted with miR-584-5p-mimics or negative control with pGL3-WWP1-WT or pGL3-WWP1-MUT. h. The expression level of WWP1 protein in GC cells after miR-584-5p expression alteration by Western blot. i. The expression level of WWP1 mRNA in GC cells after miR-584-5p expression alteration by qRT-PCR. *p < 0.05, **p < 0.01, ***p < 0.001. The data expressed as the mean ± SD