Fig. 2

RSPO3 loss modestly decreases proliferation but significantly increases invasion. DU145 or 22RV1 cells were transiently transfected with control (siCtrl) or RSPO3-specific siRNA (siRSPO3). All figures shown are representative of 3 independent experiments. a qPCR and ELISA were performed for RSPO3 expression (qPCR: DU145—left, 22RV1—middle; ELISA: 22RV1—right). b 4 day proliferation assays were performed in transfected DU145 cells. Means and standard errors are shown. (*p < 0.05). c Clonogenic survival curves for radiation resistance. Surviving fraction is shown, and curves were generated using the formula: S = e (− αD − βD²). S, surviving fraction; α and β, inactivation constants; D, dose in Gy. (n.s.—not significant). d Apoptosis assay. Cells were incubated with propidium iodide and an anti-Annexin V antibody conjugated to Cy5 fluorophore, followed by flow cytometric analysis. Cells were first gated according to forward and side-scatter parameters, and fluorescence of the gated cells is shown in a 2-dimensional pseudocolour plot. Compensation controls were generated using FlowJo software using single fluorophore-stained samples. Proportion of cells in all four quadrants of the plot is given