Imaging Genome-Edited NMuMG Cells

NMuMG cells (ATCC) and NMuMG genome-modified cells⁴² (link) were grown in DMEM containing 4.5 μg/ml glucose with pyruvate, supplemented with 10% FBS and PenStrep. Cells were seeded onto Ibidi glass bottom dishes and transfected using NMuMG-optimized GeneJet transfection reagent (SignaGen) according to the manufacturer’s instructions. 24-48 h after transfection cells were imaged in an environmental control chamber at 37 C with 5% CO₂ using a Zeiss LSM880 Airyscan microscope with a 60x oil objective NA1.4.

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Maib H, & Murray D.H. (2022). A mechanism for exocyst-mediated tethering via Arf6 and PIP5K1C-driven phosphoinositide conversion. Current Biology, 32(13), 2821-2833.e6.

Publication 2022

GeneJet transfection reagent LSM880 Airyscan microscope

Cells Dishes Genome Glucose Microscope Pyruvate Transfection

Corresponding Organization : University of Dundee

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Variable analysis

independent variables

Genome modification of NMuMG cells

dependent variables

Cell imaging outcomes

control variables

Cell culture conditions (DMEM with 4.5 μg/ml glucose, 10% FBS, PenStrep)
Transfection protocol (NMuMG-optimized GeneJet)
Imaging conditions (37°C, 5% CO2, Zeiss LSM880 Airyscan microscope, 60x oil objective NA1.4)

controls

Wild-type NMuMG cells (no genome modification) as a control group

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