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. 2020 Oct 5;11:4980. doi: 10.1038/s41467-020-18735-8

Fig. 1. c-Myc activates the pan-cancer lncRNA MILIP to repress p53.

Fig. 1

a c-Myc bound to MILIP promoter in A549 and MCF-7 cells. An E-box motif, not associated with MYC target genes on Chr22, was used as a negative control. Data shown represent three independent experiments. ChIP, chromatin immunoprecipitation. b c-Myc silencing downregulated MILIP expression in A549 and MCF-7 cells. c-Myc responsive lncRNA OVAAL was used as a positive control. Data are mean ± s.d.; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparisons test. c c-Myc silencing reduced the activity of reporters with intact c-Myc binding region (BR) of MILIP promoter but not that with the c-Myc-BR deleted (ΔBR) in A549 and MCF-7 cells. Data are mean ± s.d.; n = 3 independent experiments, two-tailed Student’s t test. d Representative microscopic photographs of in situ hybridization (ISH) analysis of MILIP expression in formalin-fixed paraffin-embedded (FFPE) lung adenocarcinoma (LUAD; n = 88 biologically independent samples) compared with paired adjacent normal tissues. Scale bar, 5 µm. e Quantitation of MILIP expression as detected in (d) in FFPE LUAD in comparison with paired adjacent normal tissues. RS, reactive score. Two-tailed Student’s t test. f Ingenuity Pathway Analysis (IPA) of RNA-seq data showing that p53 signaling was the most enriched pathway in A549 cells transfected with a MILIP siRNA (si-MILIP.2) relative to those introduced with the control siRNA. Orange bars represent pathways that were activated, and blue bars, inactivated. DEGs differentially expressed genes. g c-Myc knockdown upregulated p53 expression, which was diminished by MILIP overexpression in A549 and MCF-7 cells. Data are representatives or mean ± s.d.; n = 3 independent experiments, One-way ANOVA followed by Tukey’s multiple comparisons test. h c-Myc overexpression downregulated p53 expression, which was abolished by knockdown of MILIP in A549 and MCF-7 cells. Data shown represent three independent experiments. DOX, doxycycline, 200 ng/mL. Source data of Fig. 1a–c, e–h are provided as a Source Data file.