Fig. 4

SRPK1 is a direct target of miR-1296 in HCC cells. a miR-1296 and its putative binding sequences in the 3′-UTR of SRPK1. The mutant binding site was generated in the complementary site for the seed region of miR-1296. b HCCLM3 and Hep3B cells that were transfected with precursor miR-1296 and miR-1296 inhibitors (anti-miR-1296), respectively, were subjected to qRT-PCR for SRPK1 mRNA expression. c miR-1296 overexpression reduced the expression of SRPK1 protein in HCCLM3 cells and miR-1296 knockdown increased the level of SRPK1 protein in Hep3B cells. d miR-1296 overexpression significantly suppressed, while miR-1296 loss increased the luciferase activity that carried wild-type (wt) but not mutant (mt) 3′-UTR of SRPK1. e and f The expression of SRPK1 in miR-1296 high-expressing tumors was significantly lower than that in miR-1296 low-expressing tumors, as determined by qRT-PCR and immunoblotting. g Representative immunohistochemical staining showed a weak staining of SRPK1 in miR-1296 high-expressing HCC tissue and strong staining of SRPK1 in the miR-1296 low-expressing tumor. h An inverse correlation between the levels of miR-1296 and SRPK1 mRNA was observed in HCC tissues. *P < 0.05