Figure 6. Analysis of Taspase1-mediated cleavage of in silico predicted targets.

A. HeLa cells were transfected with biosensors harboring full length TFIIA (TS-TF2A), the predicted cleavage site from DPOLZ (TS-DPOLZ), PTRZ (TS-PTRZ), or FRM4B (TS-FRM4B) together with the Tasp-mCh or inactive Tasp^T234V-mCh expression plasmid. 24 h later, trans-cleavage resulting in nuclear translocation of the biosensors was analyzed in at least 200 fluorescent living cells. Representative examples are shown. Productive cleavage was confirmed for all targets. Scale bars, 10 µm. B. Processing of full length TFIIA-GFP upon coexpression of Taspase1 but not of the inactive Tasp^T234V mutant shown by immunoblot analysis of transfected 293T cell lysates. Protein expression and cleavage was visualized by α-GFP and α-Taspase1 Ab. GAPDH served as loading control.