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. 2006 Sep;26(18):6748–6761. doi: 10.1128/MCB.00560-06

FIG. 10.

FIG. 10.

Activation of LHR gene expression in MCF-7 cells by TSA involves PKCζ/PI3K, Sp1phosphorylation, and p107 release. (A) Reporter gene analyses of LHR gene promoter activity in MCF-7 cells which were treated with or without TSA (500 μg/ml, 24 h) in the presence or absence of PS-PKCζ, wortmannin, or LY294002. Relative luciferase activities are expressed as n-fold induction of enzyme activity in the presence of TSA over that observed in the absence of TSA. (B) Reporter gene analyses of LHR gene promoter activity in MCF-7 cells cotransfected with PKCζ siRNA or a negative control (NTC). Cells were treated with or without TSA for 24 h. The PKCζ and actin protein levels in these cells are also shown. (C) Sp1 phosphorylation status in MCF-7 cells. Nuclear extracts were isolated from untreated MCF-7 cells or cells treated with TSA in the absence or presence of Gö6976, Rottlerin, PS-PKCζ, or wortmannin. Nuclear extracts were immunoprecipitated with Sp1 antibody, followed by immunoblotting (IB) with p-Ser antibody. (D) Quantitative ChIP analyses of the association of indicated transcription factors to the LHR gene promoter in MCF-7 cells. Cells were treated with or without TSA in the presence of PS-PKCζ.