Fig. 7

NLK regulated the STAT3 and EIF4A3 accelerated the biogenesis of circ_0009092 in CRC cells.AWestern blot analysis of p53, STAT3, ERK1/2 signaling pathway in CRC cells transfected with OE-circ_0009092 or SH-circ_0009092.BWestern blot analysis of CRC cells transfected with SH-circ_0009092, miR-665 inhibitor, and SH-circ_0009092+miR-665 inhibitor.CWestern blot analysis of CRC cells transfected with SI-NLK, miR-665 inhibitor, and SI-NLK+miR-665 inhibitor.DCoIP experiment of endogenous NLK and STAT3 in Lovo and HCT-116 cells.EInteraction between STAT3 and OGT/O-GlcNAcylation modification was evaluated by coIP assay in CRC cells transfected with OE-circ_0009092 or SH-circ_0009092, WCL indicates whole cell lysates.F-GThe relative CCL2 promoter transcriptional activity was detected in NLK, STAT3, miR-665 transfected CRC cells by dual luciferase assay and normalized to Renilla luciferase activity.HThe putative binding sites of EIF4A3 in the upstream region of the circ_0009092 predicted with circinteractome database.IqRT-PCR analysis of circ_0009092 expression in EIF4A3-overexpressed and EIF4A3-knockdown CRC cells.JGEPIA database was used to evaluate the expression of EIF4A3 in 275 tumor tissues and 349 normal tissues of CRC.KqRT-PCR analysis of EIF4A3 in 20 paired CRC tissues.LThe correlation between EIF4A3 and circ_0009092 expression in 20 paired CRC tissues.MRIP assay analysis of interaction of EIF4A3 with circ_0009092 in CRC cells. IgG was used as negative control. Statistical analysis between two groups was performed using two-tailed t-test. One-way ANOVA statistical tests were adopted for more than two groups. Data are the means ± SD of three experiments. *P< 0.05, **P< 0.01, ***P< 0.001