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. Author manuscript; available in PMC: 2014 Oct 1.
Published in final edited form as: J Biomol Screen. 2013 Jun 3;18(9):10.1177/1087057113491827. doi: 10.1177/1087057113491827

Figure 2. Senescence screen setup and analysis.

Figure 2

Primary fibroblasts (IMR90) were transduced with a H-RasG12V encoding retrovirus over two days (Day -1 and 0). Cells were then selected with puromycin (1 μg/mL) for two days, plated into 96-well plates (1,000 cells/well), and at the end of the second day treated with the kinase inhibitor library at a uniform concentration (250nM). Cells were incubated for seven additional days to allow for senescence and then fixed in a formaldehyde/glutaraldehyde solution. To visualize the SA-β-gal activity and determine cell number, fixed cells were incubated with X-gal and labeled with DAPI.