Heteroplasmic MEFs harboring the tRNAAla point mutation were transfected with GenJet DNA In Vitro Transfection Reagent version II (SL100489; SignaGen Laboratories) as suggested by the manufacturers. After 24 h, cells were sorted using a FACSAria IIU (BD FACSDiva software, Version V8.0.2) by gating on single-cell fluorescence using a 561-nm laser and 600LP, 610/20 filter set for mCherry and a 488-nm laser and 505LP, 530/30 filter set for eGFP. Sorted cells expressing mCherry and eGFP were termed “Yellow”3 (link),4 (link). Sorted cells lacking fluorescence were termed “Black”.
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Sorting Heteroplasmic MEFs with Fluorescence
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Corresponding Organization : University of Miami
Other organizations : Max Planck Institute for Biology
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Variable analysis
independent variables
- Transfection with GenJet DNA In Vitro Transfection Reagent version II
- Fluorescence expression (mCherry and eGFP)
- Cell sorting (into "Yellow" and "Black" populations)
- Cell line (Heteroplasmic MEFs harboring the tRNA^Ala^ point mutation)
- Transfection protocol (as suggested by the manufacturers)
- Positive control: Cells expressing both mCherry and eGFP, termed "Yellow"
- Negative control: Cells lacking fluorescence, termed "Black"
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