Fig. 6

KLF5 is directly targeted by miR-217-5p. a Venn diagram showed the potential target genes of miR-217-5p predicted by miR-217-5p targeted genes among microT, miRDB, miRMAP, and Targetscan. b According to the TCGA database, KLF5 was upregulated in BC patients with metastasis and predicted a poor prognosis. c and d The relative expression of KLF5 were analyzed by RT-qPCR and western blot after transfection with miR-217-5p mimics and inhibitors. e Schematic illustration of KLF5 3’UTR wild type (WT) and mutant type (MT) were displayed. f The dual-luciferase assays indicated that miR-217-5p directly bind to the 3’UTR of KLF5 and inhibit the activity of luciferase. g and h Knockdown or overexpression circROBO1 decreased or increased the mRNA and protein level of KLF5 analyzed by RT-qPCR and western blot. i-k The mRNA and protein expression level of KLF5 were detected by RT-qPCR, western blot, and immunofluorescence (IF) (Scale bar=20μm) after co-transfection with indicated vectors, siRNAs, mimics or inhibitors. l and m IF and immunohistochemistry (IHC) were performed in human liver metastatic specimens. n circROBO1 activated the pathway of mTOR, phosphorylated mTOR, PI3K 110β, AKT, and phosphorylated AKT verified by western blot after co-transfection with indicated vectors, siRNAs, mimics or inhibitors. The data are showed as the mean ± SD, *P < 0.05 **P < 0.01, ***P < 0.001 and all above experiments have been repeated for three times