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. Author manuscript; available in PMC: 2015 Dec 22.
Published in final edited form as: Dev Cell. 2014 Dec 11;31(6):722–733. doi: 10.1016/j.devcel.2014.11.012

Figure 1. Characterization of p16-3MR Cells.

Figure 1

(A) Schematic of the p16-3MR transgene. See Results and Supplemental Experimental Procedures for details.

(B) qRT-PCR analysis of RNA isolated from nonsenescent (NS) and senescent (SEN; induced by 10 Gy IR) MEFs derived from p16-3MR embryos. RNA was analyzed for mRNA levels of the indicated genes relative to actin mRNA (control for cDNA quantity) (n = 3 independent experiments; A.U., arbitrary units).

(C) Immunofluorescence of cells described in (B). Blue shows DAPI-stained nuclei; red shows mRFP immunostaining.

(D) Luminescence measurements of cells described in (B). Cells were incubated with coelentarazine and lysed, and luminescence intensity was quantified using a luminometer (n = 3; A.U., arbitrary units).

(E) WT or p16-3MR MEFs were treated with GCV (10 μg/ml) for 7 days and evaluated for the percentage of surviving cells using the MTS assay (n = 4).

Data shown are the mean ± SD. **p < 0.01, ***p < 0.001.