Culturing HEK293T and MEF Cells

HEK293T cell (ATCC) and MEF were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) with 10% fetal calf serum (Greiner Bio-one, Longwood, FL, USA) in a 37°C incubator with 5% CO₂. Cells were transfected using Lipofectamine (Invitrogen) according to the manufacturer’s instruction or using polyethylenimine (Invitrogen) following manufacturer’s protocol. Bortezomib, MG132, E64, pepstatin A, Balfilomycin A 1 and cycloheximide were purchased from Sigma. siRNA against Ltn1 was bought from Ambion. siRNA against FOXO1 were obtained from Dharmacon and have been described previously (45 (link)).

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Furtado G.V., Yang J., Wu D., Papagiannopoulos C.I., Terpstra H.M., Kuiper E.F., Krauss S., Zhu W.G., Kampinga H.H, & Bergink S. (2021). FOXO1 controls protein synthesis and transcript abundance of mutant polyglutamine proteins, preventing protein aggregation. Human Molecular Genetics, 30(11), 996-1005.

Publication 2021

HEK293T cell Dulbecco’s modified Eagle’s medium (DMEM, fetal calf serum Lipofectamine polyethylenimine Bortezomib MG132 pepstatin A cycloheximide

Bortezomib Cells Cultured medium Cycloheximide Eagle Fetal calf serum Lipofectamine Mg132 Pepstatin Polyethylenimine Sirna

Corresponding Organization : University of Groningen

Other organizations : University of Siegen, Shenzhen University

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Variable analysis

independent variables

Transfection method (Lipofectamine or polyethylenimine)
SiRNA against Ltn1
SiRNA against FOXO1

dependent variables

Not explicitly mentioned

control variables

Cell lines (HEK293T and MEF)
Culture medium (DMEM with 10% fetal calf serum)
Incubator conditions (37°C, 5% CO2)
Bortezomib, MG132, E64, pepstatin A, Bafilomycin A 1, and cycloheximide

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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