BV2 cells (ATCC, Manassas, VA, USA, Cat# CRL-2467, RRID: CVCL_5744) were cultured with high-glucose Dulbecco’s Modified Eagle Medium (DMEM; Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific) and high glucose DMEM/F12 (Thermo Fisher Scientific) containing 10% FBS at 37°C in 5% CO2 under constant temperature and humidity. After transduction with BRAFV600E, BRAFWT, and vector lentivirus, respectively, for 24 hours, cells were cultured for 24 hours in DMEM/F12 without fetal bovine serum (FBS), and then medium and cells were collected for subsequent experiments. Constructs used were: pMD2.G (Addgene, Cat# 12259, RRID: Addgene_12259), psPAX2 (Addgene, Cat# 12260, RRID: Addgene_12260), pHAGE-BRAFV600E plasmid (Addgene, Cat# 116204, RRID: Addgene_116204), pHAGE-BRAFWT plasmid (Addgene, Cat# 116719, RRID: Addgene_116719), pBabe-Puro-BRAFV600E plasmid (Addgene, Cat# 15269, RRID: Addgene_15269), gag/pol-Retroviral plasmid (Addgene, Cat# 14887, RRID: Addgene_14887), Control siRNA (Cell Signaling Technology, Boston, MA, USA, Cat# 6568S), SAPK/JNK siRNA (Cell Signaling Technology, Cat# 6232S), ERK1 siRNA (Cell Signaling Technology, Cat# 6436S), ERK2 siRNA (Cell Signaling Technology, Cat# 6578S).
Erk1 sirna
Manufactured by Cell Signaling Technology
Sourced in United States
ERK1 siRNA is a laboratory product designed to target and silence the expression of the extracellular signal-regulated kinase 1 (ERK1) gene. It is intended for use in cell-based research applications to study the role of ERK1 in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Erk2 sirna, by Cell Signaling Technology (6 mentions)
Sapk jnk sirna, by Cell Signaling Technology (6 mentions)
Bv2 cells, by American Type Culture Collection (3 mentions)
Control sirna, by Cell Signaling Technology (3 mentions)
Phage braf v600e plasmid, by Addgene (3 mentions)
Phage braf wt plasmid, by Addgene (3 mentions)
Gag pol retroviral plasmid, by Addgene (3 mentions)
Pbabe puro braf v600e plasmid, by Addgene (3 mentions)
Pspax2, by Addgene (3 mentions)
Pmd2 g, by Addgene (3 mentions)
High glucose dmem f12, by Thermo Fisher Scientific (3 mentions)
High glucose dmem, by Thermo Fisher Scientific (2 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (2 mentions)
Opti mem medium, by Thermo Fisher Scientific (1 mentions)
Jnk sirna, by Thermo Fisher Scientific (1 mentions)
6 protocols using erk1 sirna
1
BV2 Cell Culture and BRAF Transduction
2
Silencing ERK1, ERK2, and JNK in Neurons
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Primary neurons and microglial cells were seeded in 6-well plates and transfected with ERK1 siRNA (Cell Signaling Technology), ERK2 siRNA (Cell Signaling Technology), SAPK/JNK siRNA (Cell Signaling Technology) at 80% cell confluence on day 4 in culture. ERK1 siRNA, ERK2 siRNA, JNK siRNA, and Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher Scientific) were diluted in OPTI-MEM medium (Thermo Fisher Scientific), mixed gently, and incubated for 5 minutes to allow complex formation. The cells were transfected by adding the RNAi-Lipofectamine complex (Invitrogen) dropwise to medium to achieve a siRNA concentration of 100 nM. The cells were collected 48 hours after the transfection for subsequent experiments (Kawakami et al., 2016).
3
Knockdown of ERK and JNK Signaling
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Primary neurons and microglial cells were seeded in 6-well plates and transfected with ERK1 siRNA (Cell Signaling Technology, USA), ERK2 siRNA (Cell Signaling Technology), SAPK/JNK siRNA (Cell Signaling Technology) at 80% cell con uence on day 4 in culture. ERK1 siRNA, ERK2 siRNA, JNK siRNA and Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher Scienti c) were diluted in OPTI-MEM medium (Thermo Fisher Scienti c), mixed gently, and incubated for 5 min to allow complex formation. The cells were transfected by adding the RNAi-Lipofectamine complex dropwise to medium to achieve a siRNA concentration of 100 nmol/l. The cells were collected 48 h after the transfection for subsequent experiments [22] .
4
Lentiviral Transduction of BRAF Variants
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Constructs used were: pMD2.G (Addgene, Cat# 12259, USA), psPAX2 (Addgene, Cat# 12260, USA), pHAGE-BRAF V600E plasmid (Addgene, Cat# 116204, USA), pHAGE-BRAF WT plasmid (Addgene, Cat# 116719, USA), pBabe-Puro-BRAF V600E plasmid (Addgene, Cat# 15269, USA) , gag/pol-Retroviral plasmid (Addgene, Cat# 14887, USA), Control siRNA (Cell Signaling Technology, Cat# 6568S, USA), SAPK/JNK siRNA (Cell Signaling Technology, Cat# 6232S, USA), ERK1 siRNA (Cell Signaling Technology, Cat# 6436S, USA), ERK2 siRNA (Cell Signaling Technology, Cat# 6578S, USA). Microglial cell line BV2 cells BV2 cells (ATCC, Cat# CRL-2467, USA) were cultured with high-glucose DMEM (Gibco) containing 10% FBS (Gibco) and high glucose DMEM/F-12 (Gibco) containing 10% FBS at 37°C in 5% CO2 under constant temperature and humidity. After transduction with BRAF V600E , BRAF WT , and vector lentivirus, respectively, for 24 h, cells were cultured for 24 h in DMEM/F12 without FBS, and then medium and cells were collected for subsequent experiments.
5
Knockdown of ERK and JNK Signaling
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+ Expand
Primary neurons and microglial cells were seeded in 6-well plates and transfected with ERK1 siRNA (Cell Signaling Technology, USA), ERK2 siRNA (Cell Signaling Technology), SAPK/JNK siRNA (Cell Signaling Technology) at 80% cell con uence on day 4 in culture. ERK1 siRNA, ERK2 siRNA, JNK siRNA and Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher Scienti c) were diluted in OPTI-MEM medium (Thermo Fisher Scienti c), mixed gently, and incubated for 5 min to allow complex formation. The cells were transfected by adding the RNAi-Lipofectamine complex dropwise to medium to achieve a siRNA concentration of 100 nmol/l. The cells were collected 48 h after the transfection for subsequent experiments [22] .
6
Lentiviral Transduction of BRAF Variants
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+ Expand
Constructs used were: pMD2.G (Addgene, Cat# 12259, USA), psPAX2 (Addgene, Cat# 12260, USA), pHAGE-BRAF V600E plasmid (Addgene, Cat# 116204, USA), pHAGE-BRAF WT plasmid (Addgene, Cat# 116719, USA), pBabe-Puro-BRAF V600E plasmid (Addgene, Cat# 15269, USA) , gag/pol-Retroviral plasmid (Addgene, Cat# 14887, USA), Control siRNA (Cell Signaling Technology, Cat# 6568S, USA), SAPK/JNK siRNA (Cell Signaling Technology, Cat# 6232S, USA), ERK1 siRNA (Cell Signaling Technology, Cat# 6436S, USA), ERK2 siRNA (Cell Signaling Technology, Cat# 6578S, USA). Microglial cell line BV2 cells BV2 cells (ATCC, Cat# CRL-2467, USA) were cultured with high-glucose DMEM (Gibco) containing 10% FBS (Gibco) and high glucose DMEM/F-12 (Gibco) containing 10% FBS at 37°C in 5% CO2 under constant temperature and humidity. After transduction with BRAF V600E , BRAF WT , and vector lentivirus, respectively, for 24 h, cells were cultured for 24 h in DMEM/F12 without FBS, and then medium and cells were collected for subsequent experiments.
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