Skip to main content
. 2016 Jul 29;7(34):54503–54514. doi: 10.18632/oncotarget.10950

Figure 3. TAp73β-mediated sensitivity of human Bone Sarcomas: regulation of apoptosis and clonogenicity.

Figure 3

a. Expression of TAp73β was evaluated by qRT-PCR in the RDES Ewing Sarcoma cell line, after infecting the cells with viral-supernatant of GFPsi -or TAp73si-transduced HEK293FT cells. Glyceraldehyde-3-phosphate dehydrogenase and β2-microglobulin were used as housekeeping genes. b. The basal apoptosis level was then evaluated by dosage of the caspase 3/7 activity in protein extracts from the same cells and in the same conditions as in (a). Error bars show the standard deviation for n = 3 measurements from representative experiments. c. The basal clonogenic capabilities of the cells were evaluated in the same cells and in the same conditions as in (a). One thousand cells were seeded in 6-wells plates and incubated until the possibility of macroscopic clones counting. The cells were then fixed in glutaraldehyde and stained with Crystal Violet. Error bars show the standard deviation for n = 3 measurements from representative experiments. Representative pictures of the wells in each condition were chosen. An unpaired Student's t-test was used to compare the different conditions in the caspase 3/7 activity assays and in the clonogenic assays. d. The same cells in the same conditions as in (a) were cultured for 48 h in the presence of Cisplatin at the indicated concentrations and cell viability was determined by WST-1 assay. The viability of the non-treated control of each cell line was assigned as 100%. A two-way ANOVA test was used to compare the different conditions in the viability assays.