CELL LINES
Contributor Information
- Name Matthias Hentze
- Institute European Molecular Biology Laboratory (EMBL)
Tool Details
- Tool name: HeLa Flp-In SLC25A6
- Tool type: Cell Lines
- Parental cell line: HeLa FLP-IN T-REx
- Model: Knock-Out
- Application: YBX3
- Description: To identify genes whose expression is controlled by YBX3
- Research area: Metabolism
- Production details: Adherent HeLa cells were grown at 37ÄÂĂ°C and 5% CO2 in DMEM, supplemented with 10% FBS Gold (PAA), 1% Penicillin/Streptomycin (Sigma) and 1% L-glutamine (GIBCO). Cells were grown until 70%ÄËĂÂĂÂ80% confluent, trypsinized, counted (Biorad TC20) and 200,000 cells were seeded per well in a 6-well dish for reverse transfection of siRNAs following the manufacturerÄËĂÂĂÂs recommendations (Lipofectamine RNAiMax, Invitrogen, cat# 137780755). YBX3 and control siRNAs were obtained from Dharmacon (YBX3 pool: SMARTpool ON-Targetplus L-015793-00 or YBX3 3ÄËĂÂĂ UTR targeting: ON-Targetplus L-015793-07; and control pool: ON-Targetplus non-targeting siRNAs 1-4). The siRNA used to target the YBX3s 3ÄËĂÂĂ UTR was YBX3 smartpool J-015793-07. Cells were harvested 48 hours after transfections.
- For Research Use Only
Target Details
- Target: YBX3
Application Details
- Application: YBX3
Handling
- Format: Frozen
- Growth medium: Adherent HeLa cells were grown at 37ÄÂĂ°C and 5% CO2 in DMEM, supplemented with 10% FBS Gold (PAA), 1% Penicillin/Streptomycin (Sigma) and 1% L-glutamine (GIBCO) Grown in medium containing blasticidine (5mg/ml) and hygromycin (200mg/ml). Cells were grown until 70%ÄËĂÂĂÂ80% confluent, trypsinized, counted (Biorad TC20) and 200,000 cells were seeded per well in a 6-well dish for reverse transfection of siRNAs following the manufacturerÄËĂÂĂÂs recommendations (Lipofectamine RNAiMax, Invitrogen, cat# 137780755). YBX3 and control siRNAs were obtained from Dharmacon (YBX3 pool: SMARTpool ON-Targetplus L-015793-00 or YBX3 3ÄËĂÂĂ UTR targeting: ON-Targetplus L-015793-07; and control pool: ON-Targetplus non-targeting siRNAs 1-4). The siRNA used to target the YBX3s 3ÄËĂÂĂ UTR was YBX3 smartpool J-015793-07. Cells were harvested 48 hours after transfections.
Documentation
- Available on request
Related Tools
References
- • The RNA-Binding Protein YBX3 Controls Amino Acid Levels by Regulating SLC mRNA Abundance
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