Mitochondrial Fatty Acid Synthesis Knockdown
Corresponding Organization : Center for Human Genetics
Other organizations : Children's Hospital of Philadelphia, Metabolon (United States)
Variable analysis
- Knockdown of the mtFASII pathway using Qiagen Flexitube siRNAs specific for the gene for ACP (NDUFAB1)
- Knockdown efficiency measured after 96 h using real time quantitative RT-PCR
- Cell viability as determined by cell count and trypan blue uptake
- Cell culture maintained at 37°C and 5% CO2
- HeLa cells plated in 10-cm cell culture dishes (CellTreat) at a density of 1.5 x 10^5 cells/mL in DMEM + 10% FBS (Mediatech, Gibco)
- Cells transfected with Allstars negative control siRNA (Qiagen)
- Cells transfected with siRNAs specific for METTL9, a nuclear gene encoding a mitochondrial protein
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