Skip to main content
. 2015 Dec 7;7(3):3461–3476. doi: 10.18632/oncotarget.6505

Figure 4. Mutation of predicted Zn binding regions in Casp3 preprotein abolishes Zn reversal of apoptosis induced by B-PAC-1 in MEF cells.

Figure 4

A. Nucleotide and amino acid sequence analysis showing WT and mutated Zn binding regions of Casp3 protein. B. Annexin V-PI FACS analysis showing cell death induced by B-PAC-1 (15 μM for 24 hr) with or without Zn (90 μM for 24 hr) in vector control Casp3–7 DKO, WT Casp3 or in MEFs stably transected with Casp3 constructs harboring mutated Zn binding amino acids. C. MEFs were treated as in B and mean (n = 5; ± SE) values relative to DMSO control are plotted. *Significant difference from control. D. Immunoblot analysis of total protein isolated from indicated cells showing cleavage of Casp3; (arrow indicates cleaved Casp3 band), XIAP, and full length PARP. GAPDH (250 μg from similar extracts) was probed for loading control. E. Experiment was conducted as in D showing mean (n = 3 ± SEM) densitometry analysis of cleaved Casp3 (p17) and XIAP ratios compared to WT Casp3 transfected MEF cells. *Significant difference from control.