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. Author manuscript; available in PMC: 2014 Jul 1.
Published in final edited form as: Mol Cancer Res. 2013 Apr 24;11(7):793–807. doi: 10.1158/1541-7786.MCR-12-0600

Figure 4. Downregulation of S1PR1 inhibits RH30 cell growth in vivo and in metastasis.

Figure 4

Panel A - qRT-PCR for S1PR1 in RH30 cells transfected with plasmids encoding shRNA against Renilla (control) and against S1PR1. The experiment was repeated twice on three different batches of cells with similar results. **p < 0.01. Panel B – FACS analysis of S1PR1 expression in RH30-shRenilla and RH30-shS1PR1 cells. The experiment was repeated three times with similar results. **p < 0.01. Panel C - Chemotaxis of RH30 cells transfected with plasmid encoding shRNA against Renilla (control) and against S1PR1 across Transwell membranes in response to S1P (1 μM). The results from three independent experiments are shown as means ± SD. *p < 0.05 or **p < 0.01 compared with the control. Panel D - RH30 tumor formation after downregulating S1PR1 expression. Tumor formation by RH30-shRenilla (control), and RH30-shS1PR1 cells inoculated into the hind limb muscles of SCID/Beige inbred mice. Five weeks later, mice were sacrificed and femora harvested to evaluate the size of the growing tumor. **p<0.01 compared with the control. Panel E - Detection of human RMS cells in bone marrow (BM), liver, lung, and peripheral blood (PB) by qRT-PCR. The results are shown as means ± SD. *p < 0.05 or **p < 0.01.