Fig. 6

KLF5 is an distinct target of miR-217-5p. a Left, The Venn diagram revealed the potential targeted gene of miR-217-5p among miRDB, microT, Targetscan, and miRMAP four databases; Right, the flow chart to filter for target gene of miR-217-5p step by step.. b RNA pull down assays were performed by biotin-miR-217-5p-probe to determine enrichment of KLF5 through RT-qPCR analysis. c Through TCGA analysis by bc-GenExMiner, the KLF5 expression was upregulated in basal-like subtype of BC d and e Right, miR-217-5p expression was negative correlated with KLF5 in BC samples in transcript analysis of clinical BC samples from SYSUCC. Left, the expression of circEZH2 was positive correlated with KLF5 in clinical BC tissues from SYSUCC in mRNA level. Both were conducted by spearman correlation analysis. f and g RT-qPCR, and western blots were used to analyze KLF5 expression after co-transfection with siRNA, vector, mimic or inhibitor as indicated. h Flow diagram indicated that establishment of xenograft-induced metastatic mice model and multi-organ metastases were analyzed by western blot and rt-qPCR. I RT-qPCR was conducted to reveal the expression pattern in different metastases compared with parental cells. j and k The expression FUS and KLF5 in different metastases were determined by western blot. P = parental; LM = lung metastases; BM = brain metastases; HM = hepatic metastases. l The pearson correlation analysis was performed to identify the protein expression association between FUS and KLF5 in metastases. m The doxycycline-induced knockdown of KLF5 in circEZH2 overexpresed cells was verified by westernblot. n and o Migration Transwell rescue assay and liver metastatic mice rescue models were performed to prove the influence of KLF5 knockdown on circEZH2 overexpression cells. The scale bar of Transwell was 100µm. The liver metastases were quantitated by IVIS and number of liver metastases were marked with yellow arrows. p Immunohistochemistry (IHC) of FUS was performed in metastases of KLF5 knockdown group and its control group (Low power field: Scale bar 100 μm; High power field: Scale bar 20µm). The data was showed as the mean ± SD and all experiments were repeated at least three times, ns no significant, *P < 0.05 **P < 0.01, ***P < 0.001