HeLa (ATCC, CCL-2, RRID: CVCL0030), U2OS (ATCC HTB-96, RRID:CVCL0042), H2030 (ATCC CRL-5914, RRID:CVCL1517), and HCC827 (ATCC CRL-2868, RRID:CVCL2063) were purchased from ATCC. HEKT293 (Thermo R70007, RRID: CVCL6911) were purchased from Thermo Scientific. HeLa and U2OS cells were cultivated in Dulbecco’s Modified Eagle’s Medium (Biochrom), containing 10% fetal calf serum, 2 mM L-glutamine, and 100 U penicillin/streptomycin. H2030, HCC827: RPMI 1640 Medium, containing 10% fetal calf serum, 2 mM L-glutamine, and 100 U penicillin/streptomycin. HEK T293: DMEM GlutaMAX™ Medium, containing 10% fetal calf serum and 100 U penicillin/streptomycin. All cell lines were tested negative for mycoplasma contamination. Cell line data were collected from Cancerrxgene (Wellcome Sanger Institute) and RNA-Seq data were obtained from Klijn et al.21 (link).
For heat stress induction, cells were incubated at 44 °C with 5% CO2. Preliminary experiments in HeLa cells and U2OS cells revealed no substantial difference between 42 °C for 4 h and 44 °C for 1 h on RNA level in our hands13 (link). Thus, the latter conditions were applied for subsequent experiments, as they induced SatIII foci in a comparable or even stronger fashion.
For heat stress induction, cells were incubated at 44 °C with 5% CO2. Preliminary experiments in HeLa cells and U2OS cells revealed no substantial difference between 42 °C for 4 h and 44 °C for 1 h on RNA level in our hands13 (link). Thus, the latter conditions were applied for subsequent experiments, as they induced SatIII foci in a comparable or even stronger fashion.
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