Fig. 4.

Hes-1 acts as a repressor of Tlx3 promoter: a Schematic of Tlx3 promoter-driven d2EGFP having a half-life of 2 h so that the change in Tlx3 expression is accurately reflected by the GFP expression. b–e FACS analysis of IMR32 cells transfected with pTlx3 1310-d2GFP in the presence or absence of Hes-1 or NICD. f Graph depicting the percentage of GFP-positive cells obtained with FACS analysis. Transfection with pTlx3-1310-d2EGFP alone significantly increased (p < 0.001) the percentage of GFP-expressing cells, whereas co-transfection with Hes-1 and NICD significantly reduced (p < 0.05 and p < 0.001, respectively) the percentage of GFP-expressing cells. g Schematic of Tlx3 promoter-driven luciferase reporter system. h Luciferase assay of Tlx3 promoter showed a significant increase (p < 0.001) in activity compared to control. Expression of Hes-1 alone with Tlx3 promoter showed a significant decrease (p < 0.001) in Tlx3 promoter activity. i–j RT-PCR analysis of Tlx3 in IMR32 cells showed a reduction in Tlx3 expression upon Hes-1 transfection. Data are expressed as mean ± SD of triplicates (n = 3) from three different experiments