Fig. 4

Circ-LRIG3 forms a ternary with EZH2 and STAT3. a The interaction probability between circ-LRIG3 and EZH2 predicted by RPISeq online software based on RF and SVM classifiers (Interaction probabilities generated by RPISeq range from 0 to 1. In performance evaluation experiments, predictions with probabilities > 0.5 were considered “positive.”). b qRT-PCR analysis verifying the efficiency of circ-LRIG3 probe. c, d RNA pull-down assay using circ-LRIG3 probe coupled Western blot analysis of EZH2 protein expression. e RIP assay using anti-EZH2 antibody coupled qRT-PCR analysis of circ-LRIG3 enrichment. f Western blot analysis of the indicated protein levels in circ-LRIG3-overexpressing HepG2 cells treated with EZH2 siRNA or GSK-126. g CCK-8 and Transwell assays detecting the cell viability and invasion in circ-LRIG3-overexpressing HepG2 cells treated with EZH2 siRNA or GSK-126. h RPISeq online software predicting the interaction probability between circ-LRIG3 and STAT3. i, j RNA pull-down assay using circ-LRIG3 probe and RIP assay using anti-STAT3 antibody in HepG2 and SMMC-7721 cells. k Co-IP assay in circ-LRIG3-silenced SMMC-7721 cells using anti-EZH2 antibody, followed by Western blot analysis of EZH2 and STAT3 protein levels. l The co-location between circ-LRIG3, EZH2 and p-STAT3. *P < 0.05, **P < 0.01