Fig. 1

Oncogenic lncRNAs were revealed as Oct4 transcriptional downstream targets in lung cancer using ChIP-seq, ChIP-PCR and qRT-PCR assays. a Flowchart of strategies used to identify Oct4 targeted downstream lncRNAs. b The Oct4 binding peaks at promoter and enhancer regions of three representative lncRNAs NEAT1, MALAT1 and UCA1. c Schematic diagram depicting the ChIP-PCR primers for amplification of the regions including Oct4 binding sites around the eight lncRNAs genomic locus (indicated as blue arrows). TSS: transcription start site as indicated by (+1). Oct4 binding regions were classified as enhancer (E) or promoter (P). d ChIP-PCR analysis of Oct4 occupancy at the binding sites of the eight lncRNAs genomic loci. GAS5 serves as a negative control lncRNA. Results are normalized to input by semi-quantitative analysis. IgG serves as an experimental negative control. Data represent mean ± SEM. P-values were determined by two-way ANOVA. e, f qRT-PCR analysis of eight lncRNAs expressions in A549 cells stably overexpressing Oct4 (Oct4#1, Oct4#2) (e) or Oct4-silenced A549 cells (si-Oct4#1, si-Oct4#2) (f). GAS5 serves as a negative control lncRNA. Target lncRNA expression levels were normalized to GAPDH expression levels. Data represent mean ± SEM. P-values were determined by two-tailed Student t-test. *P < 0.05; **P < 0.01; ***P < 0.001