siRNA ATX-1 or ENPP2

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	ATX‐1; #s10267, forward: 5′‐CCAACUCACUACUACAGCAtt‐3′, reverse: 5′‐UGCUGUAGUAGUGAGUUGGaa‐3′; si‐ATX‐2; #cs10268, forward: 5′‐CACUUGUGAUGAUAAGGUAtt‐3′, reverse: 5′‐UACCUUAUCAUCACAAGUGca‐3′) Add diluted siRNA to diluted Lipofectamine Reagent Incubate for 5 min at RT and add to cells. Incubate cells for 1–3 days at 37°C.

Protocol tips
ATX‐1; #s10267, forward: 5′‐CCAACUCACUACUACAGCAtt‐3′, reverse: 5′‐UGCUGUAGUAGUGAGUUGGaa‐3′; si‐ATX‐2; #cs10268, forward: 5′‐CACUUGUGAUGAUAAGGUAtt‐3′, reverse: 5′‐UACCUUAUCAUCACAAGUGca‐3′) Add diluted siRNA to diluted Lipofectamine Reagent Incubate for 5 min at RT and add to cells. Incubate cells for 1–3 days at 37°C.

Publication protocol

For small interfering RNA (siRNA) experiments, cells were plated on 60‐mm‐diameter dishes at 70% confluence. Silencer Select Predesigned siRNA specific for human ATX (ENPP2) (si‐ATX‐1; #s10267, forward: 5′‐CCAACUCACUACUACAGCAtt‐3′, reverse: 5′‐UGCUGUAGUAGUGAGUUGGaa‐3′; si‐ATX‐2; #cs10268, forward: 5′‐CACUUGUGAUGAUAAGGUAtt‐3′, reverse: 5′‐UACCUUAUCAUCACAAGUGca‐3′) or negative control siRNA (Applied Biosystems, Foster City, CA, http://www.appliedbiosystems.com) were prepared with transfection reagent Lipofectamine 2000 according to the manufacturer's instructions.

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Manufacturer protocol

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