2 cells and lncRNA-LALR1-up-regulated CCL-9.1 cells. The TOP/FOP ratio was higher in lncRNA-LALR1-up-regulated CCL-9.1 cells and lower in lncRNA-LALR1-down-regulated BNL CL.2 cells when compared to the respective control cells (Fig. 6A). As Fig. S7A shows, the

expression levels of cyclin D1, Axin2, and TCF7 were increased with the overexpression of lncRNA-LALR1 and decreased by the knockdown of lncRNA-LALR1. We further investigated the expression levels of Wnt/β-catenin pathway components in lncRNA-LALR1-up-regulated CCL-9.1 cells and lncRNA-LALR1-down-regulated BNL CL.2 cells. First, we analyzed the expression levels of β-catenin degradation components, including selleck screening library Axin1, GSK-3β, and APC. There was a decrease in the protein and messenger RNA (mRNA) levels of Axin1 as a result of the overexpression find more of lncRNA-LALR1, and knockdown of lncRNA-LALR1 resulted in an increase in Axin1. However, no significant difference was observed in the protein and mRNA levels of GSK-3β and APC (Fig. 6B,C). Our results also showed that overexpression of lncRNA-LALR1 resulted in a decrease in

the level of phosphorylated β-catenin protein (inactive) and that the level of nonphosphorylated β-catenin protein (active) increased (Fig. 6B). As Fig. 6D shows, active β-catenin was translocated to the nucleus, and total β-catenin staining increased because of the overexpression of lncRNA-LALR1. Western blot analysis (Fig. 6E) of β-catenin demonstrated that there was no significant difference in the cytoplasm protein level of β-catenin. However, the nuclear protein level of β-catenin significantly increased with the overexpression of lncRNA-LALR1. Next, we analyzed several target genes of the Wnt/β-catenin pathway that are associated with liver regeneration. The expression MCE公司 of c-myc and cyclin D1 increased with the overexpression

of lncRNA-LALR1 and declined after knockdown of lncRNA-LALR1 (Fig. 6B). We also investigated the changes in the protein levels of Wnt/β-catenin pathway components in lncRNA-LALR1-down-regulated mouse liver at 36 hours after 2/3 PH. The trend was similar to that in lncRNA-LALR1-down-regulated BNL CL.2 cells (Fig. 6F). We transfected pcDNA3.1-Axin1 into lncRNA-LALR1-up-regulated CCL-9.1 cells and control cells, and performed a TOP/FOP ratio analysis (Fig. S7B), western blots of Wnt/β-catenin pathway components (Fig. S7C), BrdU ELISA assays (Fig. S7D), EdU immunofluorescence (Fig. S7E), and FACS analysis (Fig. S7F). These analyses indicated that overexpression of Axin1 attenuated the function of lncRNA-LALR1 which activated the Wnt/β-catenin pathway and facilitated hepatocyte proliferation and cell cycle progression.