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At 24 and 48?h post-transfection, both siRNAs reduced SphK activity by 68�C76% for siSPHK1 #1 and 86�C87% for siSPHK1 #2 compared to a scrambled siRNA control (Figure?1B; values are the averages of two independent experiments with three replicates in each experiment). To demonstrate the specificity http://www.selleckchem.com/products/Rapamycin.html of SPHK1 siRNAs, levels of SPHK1 and SPHK2 mRNA were measured at 48?h post-transfection. The data showed a 37�C72% decrease in SPHK1, but not SPHK2, expression, with siSPHK1 #2 being more effective than siSPHK #1 when compared to control buffer or scrambled siRNA-transfected cells (Figure?1B). To determine whether SPHK1 knockdown inhibited anchorage-dependent growth of melanoma cells, siRNAs targeting SPHK1 were introduced http://www.selleck.cn/products/PD-98059.html into UACC 903 or 1205 Lu cell lines and cell viability measured. Compared to control scrambled siRNA-transfected cells, a 22�C50% decrease in cell viability was observed when SPHK1 protein expression was decreased (Figure?1C, P? http://www.selleckchem.com/products/Everolimus(RAD001).html P?