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. 2017 Mar 13;8(20):32696–32705. doi: 10.18632/oncotarget.16158

Figure 4. Binding of linc00673 to EZH2 repressed HOXA5 transcription.

Figure 4

(A) The interaction probability of linc00673 with Enhancer of Zeste Homolog 2 (EZH2) was predicted by RNA-Protein interaction prediction website (http://pridb.gdcb.iastate.edu/RPISeq/). (B, C and D) mRNA and protein changes of HOXA5 in A549 and SPC-A1 when transfected with siRNA-EZH2 on qRT-PCR and Western blot. (E, F) RNA binding protein immunoprecipitation (RIP) assay with rabbit monoclonal anti-EZH2 and preimmune IgG from A549 and SPC-A1 cell extracts. Expression levels of linc00673 RNA were presented as fold enrichment in EZH2 relative to IgG immunoprecipitates on qRT-PCR analysis. (G) Western blot analysis showed that linc00673 could pull down the EZH2 protein. HuR protein acted as a positive control. (H) ChIP of EZH2 occupancy in the NCALD promoter in A549 cells. The data represent the mean ± s.d. of three independent experiments. *P < 0.05, **P < 0.01.