RNA-seq of a human mammary fibroblast cell line in proliferation and IR-induced senescence with and without IL1-alpha treatment.
RNA-seq of a human mammary fibroblast cell line in proliferation and IR-induced senescence with and without IL1-alpha treatment.
Cell monolayers were trypsinized, neutralized with medium, washed with PBS, and centrifuged. Washed cell pellets were resuspended in Macherey-Nagel Nucleospin RNA Plus Lysis Buffer. Not applicable- no dissections. DMEM + glutamine + pyruvate + pen/strep in 5% O2 + 5% CO2 incubator.  Proliferating cells were treated with 10 pg/ml Interleukin-alpha (Sigma I2778)for 24 hours. M168 cells were induced into senescence by irradiating with 40 Gy of 120 KV X-rays produced by a Faxitron 43855D generator. Cells were incubated for 10 days post irradiation before harvesting for RNA isolation. For IL1-alpha treatment of irradiated cells, the cells were irradiated with 40 Gy of 120 KV X-rays as above and cells were incubated for 9 days and then treated for 24 hours with 10 pg/ml of IL1-alpha (Sigma I2778). Data were normalized with EdgeR and Limma voom  RNA was extracted with the Macherey-Nagel Nucleospin RNA Plus. NEBNext Ultra II Directional RNA Library Kit with PolyA RNA selection