Fig. 4: NFIA was the direct target of miR-671.

A, B Identify potential miR-671 target genes by 4 common miRNA prediction databases and GSE21034. C–E The correlations between miR-671 expression and CFL2, NFIA, and RBMS3 expression. F, G CFL2, NFIA and RBMS3 mRNA and protein expressions in RWPE-1 and PCa cell lines were examined by qPCR and WB. H WB analysis revealed a negatively correlation of NFIA expression with miR-671 expression in PCa cells. RBMS3 did not accept the regulation of miR-671. I NFIA IHC staining in xenografts with LV-in-miR-671 cells or LV-in-NC cells. J-Left: potential miR-671 binding sequences in the 3’UTR of NFIA mRNAs. J-Right: luciferase activity assays showed that the overexpression of miR-671 significantly reduced the luciferase activity of binding site of NFIA, and the mutation of binding site blocked the interaction. Magnification, ×200. Scale bars, 100 μm. The data were presented as means ± SD from three biological replicates. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; Student’s t-test. N, adjacent normal tissues; P, primary localized PCa tissues; M, metastatic PCa tissues; WT, wide type; MT, mutant type.