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. 2015 Oct 26;7(11):12331–12343. doi: 10.18632/oncotarget.6329

Figure 4. Analysis of protein levels and cleavage following p73 and p53 transfection.

Figure 4

ZR75–1 was transfected with plasmids bearing different p73 isoforms or p53. Attached (A) and detached (D) cells were harvested separately and analyzed by Western blotting. A. Transfection by Flag-TAp73α or Flag-ΔNp73α. The cleavage of TAp73α or ΔNp73α was observed in detached ZR75–1 cells following transfection but there was no evidence of p53 cleavage. Interestingly, cleavage of caspases -8 and -3 was also observed. B. Analysis of p73 using antibodies recognizing either the C- or N-terminus. Two antibodies directed against the C-terminus were used: Ab-4 (for TAp73α or ΔNp73α) and an antibody, generously donated by Dr. Caput (France) for TAp73β, another apoptotic TAp73 isoform. The anti-Flag antibody recognized the N-terminus. C. Comparison of p73 cleavage in ZR75–1 and caspase-3-nul MCF-7 following TA or ΔN transfection. Neither TAp73 nor ΔNp73 was cleaved in MCF-7 after transfection. D. Transfection of the ZR75–1 cell line with p53. Neither TAp73 nor ΔNp73 was cleaved in the ZR75–1 cell line after transfection with p53. By contrast, p53 and, very interestingly, caspase 3 were cleaved in these cells.