ON-TARGETplus Mouse Sphk1 (20698) siRNA - SMARTpool

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	The ON-TARGET plus SMART-pool of double-stranded small interfering RNA directed against mouse SphK1 was from Dharmacon Thermo Scientific. Spns2 transient interference was achieved by double-stranded mouse specific small interfering RNAs 5′- GCUAGUAGGUAUUGGUGAA (dT)(dT) -3′ (siSpns2a), 5′-CUCAUCUUUGGAGCCAUUA (dT)(dT) -3′ (siSpns2b), and 5′-CCAUCUUCAUCUGCCUCAU (dT)(dT) -3′ (siSpns2c) from Sigma. S1P1 transient interference was achieved by double-stranded human specific small interfering RNAs 5′-GCAGCUCGGUCUCUGACUA (dT)(dT) -3′ (siS1P1a) and 5′- GGAGAACAUCUUUGUCUUG (dT)(dT) -3′ (siS1P1b) from Dharmacon Thermo Scientific. S1P3 transient interference was achieved by double-stranded human specific small interfering RNAs 5′- GAACGCAGCACUUCAGAAU (dT)(dT) -3′ (siS1P3a) and 5′- GGGAUGUGCUGGCUCAUUG (dT)(dT) -3′ (siS1P3b) from Dharmacon Thermo Scientific. Aleatory sequence scrambled siRNA was from Eurogentec (Angers, France). Transfections were carried out using the Lipofectamine 2000 reagent (Invitrogen) as previously reported (Brizuela et al., 2010).

Protocol tips

The ON-TARGET plus SMART-pool of double-stranded small interfering RNA directed against mouse SphK1 was from Dharmacon Thermo Scientific. Spns2 transient interference was achieved by double-stranded mouse specific small interfering RNAs 5′- GCUAGUAGGUAUUGGUGAA (dT)(dT) -3′ (siSpns2a), 5′-CUCAUCUUUGGAGCCAUUA (dT)(dT) -3′ (siSpns2b), and 5′-CCAUCUUCAUCUGCCUCAU (dT)(dT) -3′ (siSpns2c) from Sigma. S1P1 transient interference was achieved by double-stranded human specific small interfering RNAs 5′-GCAGCUCGGUCUCUGACUA (dT)(dT) -3′ (siS1P1a) and 5′- GGAGAACAUCUUUGUCUUG (dT)(dT) -3′ (siS1P1b) from Dharmacon Thermo Scientific. S1P3 transient interference was achieved by double-stranded human specific small interfering RNAs 5′- GAACGCAGCACUUCAGAAU (dT)(dT) -3′ (siS1P3a) and 5′- GGGAUGUGCUGGCUCAUUG (dT)(dT) -3′ (siS1P3b) from Dharmacon Thermo Scientific. Aleatory sequence scrambled siRNA was from Eurogentec (Angers, France). Transfections were carried out using the Lipofectamine 2000 reagent (Invitrogen) as previously reported (Brizuela et al., 2010).

Publication protocol

The ON-TARGET plus SMART-pool of double-stranded small interfering RNA directed against mouse SphK1 was from Dharmacon Thermo Scientific. Spns2 transient interference was achieved by double-stranded mouse specific small interfering RNAs 5′- GCUAGUAGGUAUUGGUGAA (dT)(dT) -3′ (siSpns2a), 5′-CUCAUCUUUGGAGCCAUUA (dT)(dT) -3′ (siSpns2b), and 5′-CCAUCUUCAUCUGCCUCAU (dT)(dT) -3′ (siSpns2c) from Sigma. S1P1 transient interference was achieved by double-stranded human specific small interfering RNAs 5′-GCAGCUCGGUCUCUGACUA (dT)(dT) -3′ (siS1P1a) and 5′- GGAGAACAUCUUUGUCUUG (dT)(dT) -3′ (siS1P1b) from Dharmacon Thermo Scientific. S1P3 transient interference was achieved by double-stranded human specific small interfering RNAs 5′- GAACGCAGCACUUCAGAAU (dT)(dT) -3′ (siS1P3a) and 5′- GGGAUGUGCUGGCUCAUUG (dT)(dT) -3′ (siS1P3b) from Dharmacon Thermo Scientific. Aleatory sequence scrambled siRNA was from Eurogentec (Angers, France). Transfections were carried out using the Lipofectamine 2000 reagent (Invitrogen) as previously reported (Brizuela et al., 2010).

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