The 2/3 PH in C57BL/6 mice caused an increase in lncRNA-LALR1 expression that was detectable at 6 hours, peaked between 18 and 24 hours, and returned to almost normal levels by 72 hours after surgery (Fig. 2A). The timing of the

lncRNA-LALR1 surge suggested that it might be involved in liver regeneration. We also analyzed lncRNA-LALR1 expression in purified hepatocytes from the mouse liver Pirfenidone cell line samples at various timepoints after depleting the nonparenchymal cells. The trend (Fig. S4E) was similar to that of the mouse liver samples at various timepoints after 2/3 PH (Fig. 2A). Next, in situ hybridization was performed to analyze lncRNA-LALR1 expression in the mouse liver samples at 0 and 18 hours after surgery (Fig. S4F). The transcript of lncRNA-LALR1 was mainly located in the nucleus and cytoplasm of hepatocytes and was up-regulated at 18 hours after surgery.

These results suggest that lncRNA-LALR1 MG-132 research buy is specifically up-regulated in hepatocytes after 2/3 PH. The full-length sequence of lncRNA-LALR1 and the transcription start and end sites are presented in Fig. S4A. Next, we detected lncRNA-LALR1 in the BNL CL.2 cells (mouse embryo liver cell line) and mouse liver samples using northern blot analysis (Fig. 2B). Our results indicated that lncRNA-LALR1 was present and that the length of the lncRNA-LALR1 fragment was similar to that determined by RACE analysis. The transcript for lncRNA-LALR1 was located both in the nucleus and in the cytoplasm of BNL CL.2 cells, and the expression of lncRNA-LALR1 in the nucleus was higher than that in the cytoplasm (Fig. 2C). The qRT-PCR analysis

revealed significantly higher lncRNA-LALR1 上海皓元 expression in BNL CL.2 cell than in CCL-9.1 cells (Fig. S4B). To investigate the biological functions of lncRNA-LALR1 in vitro, we constructed CCL-9.1 cells with stable overexpression of lncRNA-LALR1 and BNL CL.2 cells with stable down-regulation of lncRNA-LALR1 (Fig. S4C,D). To investigate the role of lncRNA-LALR1 in hepatocyte proliferation, cell counting kit-8 assays, bromodeoxyuridine (BrdU) enzyme-linked immunosorbent assay (ELISA) assays, EdU immunofluorescence, and BrdU immunocytochemistry staining were performed in the lncRNA-LALR1-down-regulated BNL CL.2 cells and the lncRNA-LALR1-up-regulated CCL-9.1 cells. The cell counting kit-8 assays (Fig. 3A) and BrdU ELISA assays (Fig. 3B) indicated that cell proliferation was reduced by the knockdown of lncRNA-LALR1 in BNL CL.2 cells and enhanced by the overexpression of lncRNA-LALR1 in CCL-9.1 cells. As Fig. 3C,D shows, lncRNA-LALR1-up-regulated CCL-9.1 cells had higher numbers of BrdU and EdU-positive nuclei than the control cells, and the number of EdU-positive nuclei was lower in lncRNA-LALR1-down-regulated BNL CL.