Silencing of endogenous RGS2 enhances oxytocin receptor signalling via p44/42 MAPK in Hs578T cells. (a) Hs578T cells transfected with either a scrambled control small interfering (si)RNA (SiCON) or an siRNA targeting RGS2 (siRGS) were serum and insulin starved for 2 hours then stimulated with 1 × 10-7 mol/l oxytocin. Cells were harvested at timepoints up to 120 minutes after oxytocin addition and lysates analysed for levels of phospho- and total p44/42 mitogen-activated protein kinase (MAPK) by immunoblotting. (b) Quantitation of siRNA knockdown from two independent transfections. The upper plot is the quantitation of the blots shown in panel a. siRGS2 caused a significant (P < 0.001) increase in phosphorylation in response to oxytocin. (c) Quantitative real-time PCR analysis of RGS2 expression in Hs578T cells transfected with the siRGS2. Each bar represents the mean ± 95% confidence limits of the fold difference in expression compared with the mean expression in the siCON transfected cells. Data from four samples harvested from two independent transfections is shown (one of which was also used in the lower oxytocin response experiment shown in panel b). Significant differences are indicated (**P < 0.01).
Smalley et al. Breast Cancer Research 2007 9:R85 doi:10.1186/bcr1834