Research Paper Volume 6, Issue 11 pp 921—930

Figure 2. Cell cycle progression and cell death by TAp73β expression. TAp73 overexpression in SaOs-2 Tet-On cell lines does not induce proliferation. (a) PARP1-cleavage (arrow) induced by TAp73β-expression confirms induction of cell death at 24h after doxycycline (2 μg/ml) addiction. (b) Cell death assessed by sub-G1 population after PI staining. Induction of cell death is evident only after 24h of TAp73 induction. Data indicate average of triplicates and standard deviation. Blue arrows indicate the time points used for metabolomics analysis (same cultures as shown here). (c) Cell cycle profile after induction of TAp73β determined by PI staining and cytofluorimetric analysis. Controls were left untreated (0h) or treated with vehicle for 72h to account for changes induced by confluence. Data indicate average of triplicates and standard deviation. Blue arrows indicate the time points used for metabolomics analysis (same cultures as shown here). (d) TAp73β and p21 expression were assessed by western blotting after treatment with doxycycline (2μg/ml) for the indicated times. TAp73 expression was detected using HA antibody to the N-terminal HA tag. Tubulin was used as loading control. Controls are as in Figure 1.