Fig. 3: The opposite targeting of p110δ and RhoA strongly affects the proliferation, apoptosis and metastasis of melanoma tumour cells.

a Cell proliferation in tumours excised from mice that were treated concomitantly (starting on day +10) with intravenous injections of WT macrophages and intratumoural injections of p190RhoGAP siRNA or intravenous injections of δ^D910A/D910A macrophages and intratumoural injections of p190RhoGAP siRNA was determined by BrdU incorporation (brown spots) (right panels). Scale bar = 50 μm. Comparison of BrdU-positive cells in tumours from mice that were treated with intravenous injections of WT macrophages and intratumoural injections of p190RhoGAP siRNA and mice treated with intravenous injections of δ^D910A/D910A macrophages and intratumoural injections of p190RhoGAP siRNA (left panel). The symbols on the different groups denote data from eight fields/section of three sections of stained cells. b Apoptosis in tumours excised from mice that were treated concomitantly (starting on day +10) with intravenous injections of WT macrophages and intratumoural injections of p190RhoGAP siRNA or intravenous injections of δ^D910A/D910A macrophages and intratumoural injections of p190RhoGAP siRNA was determined by TUNEL assay (brown spots) (right panels). Scale bar = 50 μm. Comparison of TUNEL positive cells in tumours from mice that were treated with intravenous injections of WT macrophages and intratumoural injections of p190RhoGAP siRNA and mice treated with intravenous injections of δ^D910A/D910A macrophages and intratumoural injections of p190RhoGAP siRNA (left panel). The symbols on the different groups denote data from eight fields/ section of three sections of stained cells. c Intravasation efficiency of cancer cells as determined by tumour cells blood burden at the end point of the experiment in NSG mice which received WT or δ^D910A/D910A macrophages concomitantly (starting on day +10) with intratumoural injections of p190RhoGAP siRNA. Each symbol on the different groups denotes data from a different animal of the respective group. d Invasion of cancer cells as determined by immunohistological staining of vimentin (brown) in the lungs of NSG mice which received WT or δ^D910A/D910A macrophages concomitantly (starting on day +10) with intratumoural injections of p190RhoGAP siRNA. Scale bar = 50 μm. All immunostainings were performed on tumour sections from tumours excised at the end point of each experiment. All graphs represent means ± s.e.m. Statistically significant differences are indicated by ** (P < 0.01) or *** (P < 0.001), as determined by the Mann-Whitney U test. e Graphical representation of experimental events chronologically.