Fig. 6: miR-512-5p is extremely downregulated in LUAD cells and targets IKBKBAS. | Cell Death & Disease

Fig. 6: miR-512-5p is extremely downregulated in LUAD cells and targets IKBKBAS.

From: Novel cytoplasmic lncRNA IKBKBAS promotes lung adenocarcinoma metastasis by upregulating IKKβ and consequential activation of NF-κB signaling pathway

Fig. 6

A Half-life of IKBKBAS in A549, H1299, and HCC827 cells detected by qRT-PCR. B Basal expression of miR-512-5p in A549, H1299, or HCC827 cells detected by qRT-PCR. C The effect of miR-512-5p inhibitor or mimics on expression of miR-4741, IKBKBAS, or IKKβ mRNA in A549 (left) and HCC827 (right) cells was assayed by qRT-PCR. D The effect of miR-4741 inhibitor or mimics on expression of IKKβ in A549 and HCC827 cells were assayed by western blot. E Schematic representation of the predicted binding sites for miR-512-5p on IKBKBAS and IKKβ 3′UTR, and the site mutagenesis design for the luciferase reporter assay. F Luciferase reporter assay confirmed the interaction between miR-512-5p and IKBKBAS in A549 and HCC827 cells. A549 and HCC827 cells were co-transfected with miR-512-5p mimic or miR-512-5p inhibitor and pMIR-IKBKBAS-wt/mut plasmids.miR-NC mimics or miR-NC inhibitor were also transfected as control. Transfected cells were harvested after 48 h and subjected to luciferase activity assay. G The association among IKBKBAS, miR-512-5p, and AGO2 was ascertained by RNA pull-down assay using A549 cell lysates. Left: Detection of miR-512-5p by qRT-PCR in the sample pulled down using biotinylated IKBKBAS and IKBKBAS-AS (negative control) probes; right: Detection of AGO2 by western blot in the sample pulled down using biotinylated IKBKBAS and IKBKBAS-AS (negative control) probes. Data show mean ± S.E.M., n ≥ 3, *P < 0.05, **P < 0.01, ***P < 0.001.

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