ON-TARGETplus Mouse Eif2ak3 (13666) siRNA - SMARTpool

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	All cell lines were purchased from American Type Culture Collection (Manassas, VA, USA). Mouse colon adenocarcinoma CT26 cell lines were cultured in RPMI 1640 (Gibco) supplemented with 10% heat-inactivated FBS (PAA), 2 mM GlutaMAX I CTS (Gibco) and 100 U/mL penicillin + 100 µg/mL streptomycin (Gibco). A total of 1 × 105 CT26 cells per well were plated in six-well plates and allowed to reach 50% confluence on the day of transfection. siRNA specific for mouse PERK (ON-TARGET plus Mouse Eif2ak3 siRNA SMART pool L-044901-00-0010) and an unrelated control (ON-TARGET plus Non-targeting pool D-001810-10-05) were purchased from Dharmacon. Cells were transfected with siRNA at a final concentration of 25 nM using Lipofectamine® RNAiMAX Transfection Reagent (Invitrogen) according to the manufacturer's instructions. Knockdown of PERK was confirmed by western blotting.

Protocol tips

All cell lines were purchased from American Type Culture Collection (Manassas, VA, USA). Mouse colon adenocarcinoma CT26 cell lines were cultured in RPMI 1640 (Gibco) supplemented with 10% heat-inactivated FBS (PAA), 2 mM GlutaMAX I CTS (Gibco) and 100 U/mL penicillin + 100 µg/mL streptomycin (Gibco). A total of 1 × 105 CT26 cells per well were plated in six-well plates and allowed to reach 50% confluence on the day of transfection. siRNA specific for mouse PERK (ON-TARGET plus Mouse Eif2ak3 siRNA SMART pool L-044901-00-0010) and an unrelated control (ON-TARGET plus Non-targeting pool D-001810-10-05) were purchased from Dharmacon. Cells were transfected with siRNA at a final concentration of 25 nM using Lipofectamine® RNAiMAX Transfection Reagent (Invitrogen) according to the manufacturer's instructions. Knockdown of PERK was confirmed by western blotting.

Publication protocol

All cell lines were purchased from American Type Culture Collection (Manassas, VA, USA). Mouse colon adenocarcinoma CT26 cell lines were cultured in RPMI 1640 (Gibco) supplemented with 10% heat-inactivated FBS (PAA), 2 mM GlutaMAX I CTS (Gibco) and 100 U/mL penicillin + 100 µg/mL streptomycin (Gibco). A total of 1 × 105 CT26 cells per well were plated in six-well plates and allowed to reach 50% confluence on the day of transfection. siRNA specific for mouse PERK (ON-TARGET plus Mouse Eif2ak3 siRNA SMART pool L-044901-00-0010) and an unrelated control (ON-TARGET plus Non-targeting pool D-001810-10-05) were purchased from Dharmacon. Cells were transfected with siRNA at a final concentration of 25 nM using Lipofectamine® RNAiMAX Transfection Reagent (Invitrogen) according to the manufacturer's instructions. Knockdown of PERK was confirmed by western blotting.

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