Fig. 5

LINC01123 is a direct transcriptional target of c-Myc and inversely enhances c-Myc level. a Schematic illustration of consensus c-Myc binding sites in LINC01123 gene promoter. The indicated pGL3-based luciferase reporters were constructed containing either of the two putative c-Myc binding sites or their corresponding mutant binding sites, which were shown in the open boxes. b A549 cell was transfected with 0.5 μg, 1 μg, or 2 μg pcDNA-c-Myc. H1299 cell was transfected with si-NC or si-Myc. Forty-eight hours after transfection, cell lysates and total RNA were subjected to Western blot and qRT-PCR analyses, respectively. c, d Two hundred ninety-three T cell was co-transfected with either pcDNA c-Myc or si-Myc plus the indicated reporter constructs and Renilla luciferase plasmid. Twenty-four hours after transfection, reporter activity was measured and plotted after normalizing with respect to Renilla luciferase activity. e Lysates from A549 cell were subjected to ChIP assay. ChIP products were amplified by qPCR with the indicated pairs of primers. f–h QRT-PCR, Western blot, and immunofluorescence showed that c-Myc mRNA and protein level were positively regulated by LINC01123 in A549 and H1299 cells. Data shown are mean ± SD (n = 3). (*P < 0.05, **P < 0.01, ***P < 0.001)