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Fig. 3 | Journal of Hematology & Oncology

Fig. 3

From: Up-regulated LINC01234 promotes non-small-cell lung cancer cell metastasis by activating VAV3 and repressing BTG2 expression

Fig. 3

LINC01234 interacts with RNA-binding proteins and functions as a ceRNA for miR-27b-3p and miR-340-5p in NSCLC cells. a qRT-PCR analysis of the subcellular localization of LINC01234 in A549 and SPC-A1 cells. b RIP assays showing LINC01234 co-immunoprecipitation with EZH2, LSD1, and Ago2 from A549 and SPC-A1 cells. c RNA-pull-down assays showing EZH2, LSD1 and Ago2 binding to desthiobiotinylated LINC012345 from A549 cells. Androgen receptor (AR) RNA binding to HuR was analyzed as a positive control. d Predicted miRNA-binding sites in LINC01234 identified using online bioinformatics databases (left). Validation of miRNA sponges for LINC01234 by luciferase reporter assays. Luciferase activity was normalized to Renilla activity (right). e Schematic diagram of the predicted binding sites for miR-27b-3p and miR-340-5p in LINC01234. f Validation of miR-27b-3p and miR-340-5p sponges for LINC01234 by luciferase reporter assays. Luciferase activity was normalized to Renilla activity. g Immunoprecipitation of Ago2 and qPCR of associated RNAs (LINC01234, miR-27b-3p, and miR-340-5p). h, i Relative expression of miR-340-5p and miR-27b-3p in NSCLC cells overexpressing or depleted of LINC01234. j Relative expression of LINC01234 in NSCLC cells transfected with miR-340-5p or miR-27b-3p mimics. *P < 0.05, **P < 0.01

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