Fig. 4

Lnc-GAN1 is expressed independent of its parent gene GAN. a Lnc-GAN1 expression is lower in 30 NSCLC tissues than in paired normal lung tissues, while its parent gene GAN expression is not different between the same NSCLC and paired normal lung tissues. b There is not a correlation between Lnc-GAN1 and GAN expressions in 8 cell lines. c In dual-luciferase reporter assay, A549 cells with the reporter containing 2000-bp DNA sequence upstream of lnc-GAN1 produce significantly higher Luciferase activity than A549 cells with control reporter, indicating that the 2000-bp sequence harbors promoter binding site. Luciferase activity was calculated as a firefly/renilla luciferase ratio and expressed as mean ± SD of three independent experiments. d Chromatin immunoprecipitation (ChIP) analysis on H460 and H1650 cells with Pol II antibody and the precipitated DNAs were amplified by PCR with the primers for the 2000-bp DNA sequence upstream of lnc-GAN1. H1650 cells with high expression of lnc-GAN1 show remarkably higher PCR product than H460 cells with very low expression of lnc-GAN1, suggesting that this DNA sequence has high Pol II activity in H1650 cells