Figure 8From: Integrity of the LXXLL motif in Stat6 is required for the inhibition of breast cancer cell growth and enhancement of differentiation in the context of progesterone Stat6 gene expression is induced by progesterone. Synchronized T47D cells were treated with or without progesterone (30nM) for the indicated times (A, left panel) or for 24 h (A, right panel, and B). A. Quantitative PCR analysis. mRNA levels are expressed relative to levels in vehicle-treated control cells harvested at 6 h, arbitrarily set as 1. Right panel. Cycloheximide (5 μg/ml) or actinomycin D (10 μg/ml) was added to the medium 2 h before the addition of progesterone or ethanol (vehicle). h, hour; d, day; NS, not significant; *, P ≤ 0.05; **, P ≤0.01 versus control. B. Modulation of progesterone on Stat6 was detected in T47D cells after 24 h progesterone treatment using Confocal Laser Scanning Microscope. Coverslips seeded with synchronized T47D cells treated with or without progesterone (30nM) and vehicle were triple-stained with Stat6 antibody and one secondary antibody conjugated with FITC to permit detection of Stat6 (a), phalloidin to permit detection of actin (b), and DAPI, a DNA-binding dye, to counterstain the DNA (c). In T47D cells, Stat6 accumulated in nuclear domains that colocalized with dense regions of DAPI staining (d).Back to article page