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Fig. 5 | Molecular Cancer

Fig. 5

From: Long non-coding RNA NEAT1 promotes bone metastasis of prostate cancer through N6-methyladenosine

Fig. 5

NEAT1–1 recruited CYCLINL1 and CDK19 on RUNX2 promoter via m6A site #4. a Venn diagram showing that genes mediated by NEAT1–1 knocking down overlapped with genes mediated by NEAT1–1 site#4 m6A deletion (nt3494A deletion) in P-18 cells (P = 9.3e-10, permutation test). b A diagram of CDK19 and CYCLINL1 interaction through NEAT1 on promoter of target gene. c ChIRP assay using biotin-labeled LacZ or NEAT1–1-specific DNA probes and streptavidin beads. Pulldown DNA was analyzed by real-time PCR. All data shown were mean values ± SD (error bar) from three replicates. P values were shown in the figures. d CHART-qPCR analyswas of CYCLINL1 binding at the NEAT1–1 WT and m6A-mut in P-18 NEAT1-KO cells. Immunoprecipitated RNAs were detected by real-time PCR. All data shown were mean values ± SD (error bar) from three replicates. m6A-mut was site#4 m6A deletion in NEAT-1. P values were shown in the figures. e RUNX2 expressions were measured by qRT-PCR in P-18 and P-34 primary cells. Means and standard deviations (error bar) were determined from three replicates. Error bars represent mean ± SD for triplicate experiments. m6A-mut was site#4 m6A deletion in NEAT-1. P values were shown in the figures. f Expression of RUNX2, Tubulin, Pol II Ser-2 phosphorylations and total Pol II proteins were measured by western blot in P-18 NEAT1–1-KO cells infected with control or NEAT1–1-WT, NEAT1–1-m6A-mutant (nt3494A deletion) or NEAT1–1 promoter-binding mutant (nt874–899 deletion)

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