Fig. 5.

GAS6-AS1 increases MYC target gene expression through YBX1. A Volcano plots of RNA-seq showing the change of gene expression (fold change > 1.5, P < 0.05) in U937 cells stably transfected with scramble vector or GAS6-AS1 shRNA. B Venn diagram (left panel) was applied to identify transcription factors (TFs) regulating target gene expression, based on overlapping analysis of potential TFs using ChIP-X program and YBX1-interacting proteins from BioGRID database. C Log-rank test (right panel) showing the correlation of identified TFs with overall survival of AML cases (GSE37642). N.S., nonsignificant. D Dual-luciferase assay with a reporter carrying MYC binding sites showing the relative activity of MYC in Kasumi-1 and U937 cells stably transfected with empty vector (mock), GAS6-AS1, scramble shRNA (sh-Scb), or sh-GAS6-AS1 (n = 4). E, F Dual-luciferase assay indicating the relative activity of EP300, TP53, or TFAP2A in Kasumi-1 and U937 cells stably transfected with mock, MZF1-AS1, sh-Scb, or sh-MZF1-AS1 (n = 4). G, H ChIP and qPCR (normalized to input) indicating MYC enrichment at promoters of target genes in Kasumi-1 and U937 cells stably transfected with a series of vectors. I ChIP and real-time qRT-PCR (normalized to β-actin) showing the transcript levels of target genes. J Western blot showing protein levels of target genes in U937 cells stably transfected with sh-Scb, or sh-GAS6-AS1 #1, and those co-transfected with YBX1. Data were depicted as mean ± s.e.m., *P < 0.05