Autotaxin promotes the growth and survival of EBV-infected HL cells. (A) (top left) Semiquantitative RT-PCR analysis of equivalent starting amounts of cDNA (shown here are results of 30 cycles of amplification) from untreated EBV-positive KM-H2 cells or these cells treated for 48 hours with transfection reagent alone (Ribojuice only), Ribojuice plus scrambled siRNA, or autotaxin (ATX)-specific siRNAs. A clear difference in autotaxin mRNA levels between siRNA-transfected and control cells was detectable. (bottom panel) GAPDH mRNA levels were unaffected by these treatments. (B) Immunoblotting demonstrates knockdown of autotaxin protein in EBV-positive KM-H2 cells treated with autotaxin-specific siRNAs compared with controls. (C) Treatment of EBV-infected KM-H2 cells with autotaxin-specific siRNAs resulted in significant reduction in proliferation after 48 hours (top panel) and cell viability after 72 hours (bottom panel) compared with cells treated with transfection reagent alone or with scrambled siRNA (data not shown). RJ indicates ribojuice transfection reagent. (D) Down-regulation of autotaxin expression in EBV-infected KM-H2 cells resulted in reduced generation of LPA from LPC.