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. 2009 Feb 17;37(7):2070–2086. doi: 10.1093/nar/gkp067

Figure 1.

Figure 1.

HMGB1 up-regulates activity of human topo IIα promoter. (A) Identification of elements within human topo IIα gene promoter involved in HMGB1-mediated up-regulation of the promoter. Saos-2 cells were co-transfected with the plasmid constructs containing the –557 bp human topo IIα promoter (plasmid pTIIα-557) or one of the truncated topo IIα promoter constructs linked to the luciferase reporter gene (as detailed in Materials and methods section), and plasmid encoding HMGB1 (or empty vector). Inset, over-expression of Flag-tagged HMGB1. Lane 1, cells transfected with empty vector; lane 2, cells transfected with plasmid encoding Flag-HMGB1. Immunodetection was carried out using specific α–HMGB1 antibody. (B) Effect of mutation of ICE2 on transactivation of topo IIα promoter by HMGB1. Saos-2 cells were co-transfected with the plasmids harboring the full-length topo IIα promoter (plasmids pTIIα-617 or pTIIα-617_ICE2mt) or truncated topo IIα promoter (plasmid pTIIα-142 or pTIIα-142_ICE2mt) containing the wild-type (WT) or mutated ICE2 (ICE2mt), and either empty vector or plasmid encoding HMGB1. Luciferase activity (referred as to transactivation) in panels (A) and (B) is presented relative to the luciferase activity of the topo IIα promoter in cellular lysates without HMGB1 over-expression (taken as 1). Each transfection experiment was repeated at least five times with triplicate samples. Values are presented as the mean ± 1 SD (error bars) of luciferase activities from triplicate samples in a representative experiment.