Fig 1: The expression of Glut1 was modulated by the circ_0084043/miR-31/KLF3 regulatory axis. (a) The protein level of Glut1 in A375 and A875 cells transfected with si-circ_0084043, si-NC, si-circ_0084043 + miR-31 inhibitor, or si-circ_0084043 + inhibitor NC was measured by western blot. (b) The protein level of Glut1 in A375 and A875 cells transfected with miR-31 mimic, miRNA NC, miR-31 mimic + pcDNA-KLF3, or miR-31 mimic + pcDNA-Control was measured by western blot. *P < 0.05.
Fig 2: KLF3 was targeted by miR-31, and its overexpression abolished the role of miR-31 mimic. (a) The binding site between miR-31 and KLF3 3'-UTR was forecasted by the online tool TargetScan. (b and c) The interaction between miR-31 and KLF3 was confirmed by the dual-luciferase reporter assay. (d–f) The expression of miR-31 and KLF3 was checked in the cells transfected with miR-31 mimic. (g and i) The expression of KLF3 at mRNA and protein levels in the cells transfected with pcDNA-KLF3 was examined by qRT-PCR and western blot. (h and j) Cell proliferation was assessed by the MTT assay. (k) Cell apoptosis was examined by the flow cytometry assay. (l–n) The progression of glycolysis was evaluated, according to glucose consumption, lactate production, and ATP concentration. *P < 0.05.
Fig 3: Kaplan–Meier estimates of disease-free survival of patients with colorectal cancer with different expression levels of Kruppel-like factor 3 (KLF3) mRNA levelsThe 5-year disease-free survival rate for patients in the high and low KLF3 mRNA expression groups was 76.9% and 57.5%, respectively (?2=10.085, P<0.001).
Fig 4: Circ_0084043 knockdown inhibited tumor growth in vivo. (a and b) Circ_0084043 knockdown significantly reduced the tumor volume and tumor weight. (c and d) The expression of circ_0084043 and miR-31 in removed tumor tissues was detected by qRT-PCR. (e and f) The expression of KLF3 in removed tumor tissues at both mRNA and protein levels was determined by qRT-PCR and western blot. *P < 0.05.
Fig 5: The target gene of miR-365a-3p was predicted via bioinformatics analysis. (A) miR-365a-3p target genes were predicted using three miRNA databases (TargetScan, miRDB and DIANA tools). The top 100 candidate genes from each database were used for subsequent analysis (B-M) RT-qPCR was used to determine the association between miR-365a-3p expression and mRNA expression levels of the top ranked candidate target genes, including ARRB2, CBFB, ESRRA, USP33, TBK1, SOCS5, WDR37, ZNF148, USP48 and E2F2. (N) Western blotting and RT-qPCR analysis of protein and mRNA expression levels of KLF3, respectively, in LOVO and SW480 cells transfected with a lentiviral vector carrying a miR-365a-3p-specific mimic or inhibitor, respectively. miR/miRNA, microRNA; RT-qPCR, reverse transcription-quantitative PCR; KLF3, Kruppel-like factor 3; ARRB2, arrestin ß2; CBFB, core-binding factor subunit ß; ESRRA, estrogen related receptor a; USP33, ubiquitin specific peptidase 33; TBK1, TANK binding kinase 1; SOCS5, suppressor of cytokine signaling 5; WDR37, WD repeat domain 37; ZNF148, zinc finger protein 148; USP48, ubiquitin specific peptidase 48; E2F2, E2F transcription factor 2. **P<0.05 compared with group LV-INC or LV-mNC. *** P<0.01 compared with group LV-INC or LV-mNC.
Supplier Page from Abcam for Anti-KLF3 antibody