Hela Kyoto (HK) cells were grown in Dulbecco's modified Eagle's medium (DMEM) (Sigma) supplemented with 10% fetal calf serum (FCS; Sigma) and 1% penicillin/streptomycin at 37°C and 5% CO2. Transfection was performed as described in (19 (link)) using X-tremeGene (Roche Applied Science) according to the manufacturer's instruction. Expression levels of HK cells stably transfected with eGFP and eGFP-tagged histone H3 variants and mutants were quantified using a FACSCanto machine (Becton Dickinson). Human primary mesenchymal stem cells were purified from lipoaspirates and cultured and transfected by electroporation using Nucleofector™ Technology (Lonza) as described in (20 (link)).
Protocol detail
Cell Culture and Transfection of Stem Cells
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Electroporation
Hela cells
Histone h3
Human
Mesenchymal stem cells
Penicillin
Streptomycin
Transfection
Corresponding Organization : Center for Integrated Protein Science Munich
Other organizations : University of Oslo, Max Planck Institute of Biochemistry
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Variable analysis
independent variables
- Transfection method (X-tremeGene vs. electroporation)
- Expression levels of eGFP and eGFP-tagged histone H3 variants and mutants in HK cells
- Expression levels in human primary mesenchymal stem cells
- Cell culture conditions for HK cells (DMEM, 10% FCS, 1% penicillin/streptomycin, 37°C, 5% CO2)
- Cell culture and transfection of human primary mesenchymal stem cells (as described in reference 20)
- Positive control: HK cells stably transfected with eGFP
- Negative control: Not explicitly mentioned
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