FIG 4.

LINC01006 acts as a sponge for miR-129-2-3p to upregulate CTNNB1. (A) Subcellular fractionation was performed to locate LINC01006 in LUAD cells. (B) The effect of LINC01006 downregulation on the relative expression level of CTNNB1 was evaluated by RT-qPCR analysis. (C) Four potential miRNAs binding to LINC01006 and CTNNB1 were discovered with the application of bioinformatics. (D) miR-129-2-3p was screened out as the miRNA that could bind to both LINC01006 and CTNNB1 through RNA pulldown assay. (E) The expression of miR-129-2-3p in healthy lung epithelial cell line (BEAS-2B) and LUAD cell lines (PC9, H1650, H1975, and A549) was examined. (F) RIP assay was implemented to demonstrate the coexistence of LINC01006, miR-129-2-3p, and CTNNB1 in RNA-induced silencing complexes (RISCs). (G) RIP assays researching the enrichment of LINC01006 were carried out when miR-129-2-3p was overexpressed or downregulated in LUAD cells. (H) RIP assays researching the enrichment of CTNNB1 were done when the expression of LINC01006 or miR-129-2-3p was inhibited in LUAD cells. (I) Luciferase reporter assay was carried out to verify the interaction between miR-129-2-3p and LINC01006 or CTNNB1 in HEK-293T or LUAD cells. (J) RT-qPCR was performed to detect the relative expression level of LINC01006 after either the overexpression or downregulation of miR-129-2-3p. (K) Copy numbers of LINC01006, miR-129-2-3p, and CTNNB1 in H1975 and A549 cells were measured by RT-qPCR. **, P < 0.01.