Silencer®_Skiv2l2 siRNA (m)

Protocol tips

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	Before RNAi knockdown, 3 × 105 N2A or P19 cells were plated on 35-mm plates. When cells were at ∼60% confluency, cells were transfected with the appropriate siRNA. Skiv2l2 knockdowns were performed by transfecting cells with either 67 nM Skiv2l2 Silencer siRNA ID #177475 or #177476 (Thermo Fisher #AM16704, #AM16708) or 67 nM Negative Control Silencer siRNA by Ambion (Thermo Fisher #AM4635). Delivery of siRNAs was accomplished with 1 mL of Opti-MEM (Thermo Fisher #31985) and 0.1% Lipofectamine RNAiMax by Invitrogen (Thermo Fisher #13778) for N2A cells or 0.1% Lipofectamine 3000 by Invitrogen (Thermo Fisher #L300008) for P19 cells according to the manufacturer's protocol.Cells treated with the siRNAs were grown for 48 h at 37°C, 5% CO2, and 95% humidity. The cells were then harvested following RNAi using simple pipetting.

Protocol tips

Before RNAi knockdown, 3 × 105 N2A or P19 cells were plated on 35-mm plates. When cells were at ∼60% confluency, cells were transfected with the appropriate siRNA. Skiv2l2 knockdowns were performed by transfecting cells with either 67 nM Skiv2l2 Silencer siRNA ID #177475 or #177476 (Thermo Fisher #AM16704, #AM16708) or 67 nM Negative Control Silencer siRNA by Ambion (Thermo Fisher #AM4635). Delivery of siRNAs was accomplished with 1 mL of Opti-MEM (Thermo Fisher #31985) and 0.1% Lipofectamine RNAiMax by Invitrogen (Thermo Fisher #13778) for N2A cells or 0.1% Lipofectamine 3000 by Invitrogen (Thermo Fisher #L300008) for P19 cells according to the manufacturer's protocol.Cells treated with the siRNAs were grown for 48 h at 37°C, 5% CO2, and 95% humidity. The cells were then harvested following RNAi using simple pipetting.

Publication protocol

Before RNAi knockdown, 3 × 105 N2A or P19 cells were plated on 35-mm plates. When cells were at ∼60% confluency, cells were transfected with the appropriate siRNA. Skiv2l2 knockdowns were performed by transfecting cells with either 67 nM Skiv2l2 Silencer siRNA ID #177475 or #177476 (Thermo Fisher #AM16704, #AM16708) or 67 nM Negative Control Silencer siRNA by Ambion (Thermo Fisher #AM4635). Delivery of siRNAs was accomplished with 1 mL of Opti-MEM (Thermo Fisher #31985) and 0.1% Lipofectamine RNAiMax by Invitrogen (Thermo Fisher #13778) for N2A cells or 0.1% Lipofectamine 3000 by Invitrogen (Thermo Fisher #L300008) for P19 cells according to the manufacturer's protocol.Cells treated with the siRNAs were grown for 48 h at 37°C, 5% CO2, and 95% humidity. The cells were then harvested following RNAi using simple pipetting.

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