Fig. 5

FLOT1 is a direct target of miR-608. a Representative IHC staining of FLOT1 in TMA. FLOT1 showed a membranous location. b Statistical analysis indicated that the expression levels of FLOT1 protein in BCa tissues were significantly higher than those in adjacent non-tumor tissues in TMA. c Representative IHC staining of FLOT1, p-AKT and p-FOXO3a in xenograft tumors treated by miR-608 in contrast with NC. d and e Western blot and qRT-PCR assays. The protein and mRNA levels of FLOT1 in BCa cells were negatively regulated after the up or down-regulation of miR-608 by using miR-608 mimic or inhibitor respectively. f Predicted miR-608 target sequences in the 3′-UTR of FLOT1. g Dual-luciferase reporter assay. The relative luciferase activity in HEK 293 T cells co-transfected with miR-608 mimic and Wt-2 vector was sharply inhibited, while in those cells simultaneously transfected with miR-608 mimic and Wt-1 vector or mutant vector (Mut-1 or Mut-2) the luciferase activity was unaffected. Error bars represent the S.D. from three independent experiments. *P < 0.05