Figure 2
From: Quantification of PRL/Stat5 signaling with a novel pGL4-CISH reporter
The dose response and time course of pGL4-CISH to PRL in T47D breast cancer cells. Following transfection of 1 × 105 T47D cells with the pGL4-CISH and renilla control reporters, transfectants were cultured in minimal defined medium without FBS for 24 hours followed by 24 hours of PRL stimulation prior to luminescence assay. The pGL4.10 reporter backbone was used to compensate DNA amount used for transfection. A. Luciferase luminescence in PRL-stimulated T47D cells transfected with varying concentrations of pGL4-CISH reporter DNA (PRL concentration, 100 ng/ml). B. Fold changes of normalized PRL-stimulated pGL4-CISH expression versus non-PRL-stimulated T47D transfectants as a function of transfected pGL4-CISH concentration (PRL concentration, 100 ng/ml). C. PRL dose response of expression of pGL4-CISH in T47D transfectants. T47D cells were transfected with 50ng pGL4-CISH/1 × 105 cells, arrested and treated with PRL (100 ng/ml) for 24 hours. D. Time course of pGL4-CISH expression in the presence of PRL (100 ng/ml).