Fig. 2

NEAT1 is regulated by c-Myc in K562 cells. a K562 cells were transfected with si-BCR-ABL and NEAT1 promoter constructs. Twenty-four hours after transfection, the luciferase activity was analyzed. b Luciferase reporter assays were performed with K562 cells using the indicated reporters, and the cells were treated with LY204002 and IM. c qPCR analysis of the expression of total NEAT1 in K562 cells following transfection with control siRNA and c-Myc siRNA, demonstrating marked dependence on c-Myc. d Luciferase reporter assays were performed with cells transfected with NEAT1 promoter constructs together with si-Myc or control siRNA. e Cells were transfected with NEAT1 promoter constructs (Renilla luciferase reporter construct as a control vector) and treated with DMSO or MG132 (Previous studies have demonstrated that NEAT1 is transcriptionally up-regulated by the proteasome inhibitor MG132 in Hela cells), and luciferase activity was measured after 24 h. e c-Myc ChIP followed by qPCR analyzed the occupation of c-Myc on the NEAT1 gene. The co-immunoprecipitated DNA was amplified by PCR using primer pairs spanning a 500 bp region. Primers that amplify non-c-Myc binding regions were used as negative control(primer NC). The data are shown as a percentage of the input