Fig. 3: Caspase-2-S384E mutant can still homodimerise but lacks enzymatic activity.
From: Phosphorylation by Aurora B kinase regulates caspase-2 activity and function
a Representative confocal images showing dimerisation of Casp2-S384E or CASP2-C320A by BiFC. U2OS-CASP2−/− cells were transfected with Casp2-S384E or CASP2-C320A BiFC constructs, respectively, together with pDsRED-Mito as transfection reporter and treated with DMSO or 100 nM BI2536 (BI) for 24 h. All cells were also treated with z-VAD (20 µM), pan-caspase inhibitor. DAPI (blue); pDsRED-Mito (red). Arrows indicate dimerised caspase-2 (Green). Scale bar = 20 μm. b Quantitation of relative BiFC-positive cells are indicated. *p < 0.05. c Immunoblot analysis of recombinant GST-caspase-2-WT, C320G and S384E protein expression and autoprocessing. ‘*’, truncated GST-caspase-2. d Caspase activity of recombinant GST-caspase-2-WT, C320G and S384E protein, was assessed by cleavage of VDVAD-AMC. GST protein was used as a control. Data represented as mean ± SEM from three independent experiments. **p < 0.01 compared to GST-caspase-2-WT.
