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. 2017 May 10;8:14995. doi: 10.1038/ncomms14995

Figure 3. RNA levels of H2AFJ and selected canonical H2A species.

Figure 3

RNA levels are compared in senescent or quiescent cells relative to proliferating cells. Random primers were used to prepare cDNA to quantify total RNA (a) whereas oligo dT was used to prepare cDNA to quantify polyA RNA (b). Shown are the mean and s.d. for triplicate qPCR values from single samples. A second biological replicate from total RNA is shown in Supplementary Fig. 4. RNA levels were normalized to GAPDH mRNA and to the levels of the respective histone mRNAs in proliferating fibroblasts. RNAs were extracted from proliferating WI-38hTERT-GFP-RAF-ER fibroblasts (Prolif.), the same cells induced into quiescence by serum starvation for 5 days (e-Quiescent) or 20 days (d-Quiescent), induced into senescence by the expression of a hyperactive RAF kinase for 5 days (e-SenRAF) or 20 days (d-SenRAF), or induced into senescence by treatment with etoposide for 5 days (e-SenETO) or 20 days (d-SenETO). RNAs were also extracted from non-immortalized proliferating WI-38 fibroblasts (Early Passage) and the same cells maintained in replicative senescence for 20 days (d-SenRep).