Abstract
INTRODUCTION. Hoechst 33342 (H342) and Hoechst 33258 (H258) bind to AT regions of the DNA minor groove. H342, but not H258, induces apoptosis in various cells (1). Sequence-selective DNA-binding drugs inhibit transcription factors from binding to their target sites on gene promoters in vitro (2). In vivo studies demonstrate that H342-induced apoptosis is associated with degradation of TATA box binding protein (TBP) (3) and accumulation of intracellular E2F-1 (4). Here, we determine the effect of H342 and H258 on in vivo luciferase gene expression in BC3H-1 myocytes using 4 pGL-3 vectors: Basic, Promoter, Enhancer, and Control.
Highlights
Hoechst 33342 (H342) and Hoechst 33258 (H258) bind to AT regions of the DNA minor groove
BC3H-1 myocytes were grown in MEME with 10% FBS, plated at a density of 1 x 104 cells/mL, and were cultured for 2 days to 60-80% confluence prior to transfection with a pGL-3 vector using the non-liposomal formulation FuGENETM 6 transfection reagent
Luciferase enzyme activity is dependent upon H342 concentration (Fig. 1)
Summary
Hoechst 33342 (H342) and Hoechst 33258 (H258) bind to AT regions of the DNA minor groove. BC3H-1 myocytes were grown in MEME with 10% FBS, plated at a density of 1 x 104 cells/mL, and were cultured for 2 days to 60-80% confluence prior to transfection with a pGL-3 vector using the non-liposomal formulation FuGENETM 6 transfection reagent.
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