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Moreover, in JIM-3 HMCLs, a significant cleavage of caspase3 was observed with AVE1642 alone, coinciding with moderate apoptosis and Noxa induction.
The combination induced a significant cleavage of procaspase-8 to its active fragments (p43/41) and procaspase-9 to its active fragments (p35/37) compared with cells treated with either of the two compounds alone.
Downstream effector caspase activation based on DEVDase activity using a probe containing CFP and YFP indicated a significant cleavage rate starting at 75 min over a 30 40 min period for 4C7.
Since catalytically inactive C163S caspase-3 did not show a significant cleavage during expression, we suspected that self-catalysis by the precursor proteins is responsible for cleavage of proteins V. Hence, purified precursors were treated with catalytically active caspase-3 to establish whether auto-processing by precursor proteins was eliminated by mutation.
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Interestingly, a third significant cleavage site was observed for Mut1 (band c), corresponding to a cleavage downstream from a 5'-TA sequence.
Indeed, we observed less potent cleavage of this protein by Staphopain A but at higher enzyme concentrations there was still significant cleavage.
No significant cleavage of caspase-3, an executioner caspase, or caspase-9, an initiator caspase, after treatment with VK1 was observed.
Analysis of the pro- and anti-apoptotic pathway proteins in 8505c cells treated with the combination of all three drugs: PLX4720 100 μM), LY294002 (50 μM) and lexatumumab (1000 ng/ml) in 8505c cells showed: (a) Activation of all extrinsic and intrinsic pathways with significant cleavage of caspase-8/-9/-3 and PARP.
Whereas the a strand was cleaved almost to completion within the first minute, significant cleavage of the b strand only became apparent after 2 min, proceeding to completion in ∼40 min.
The two female backup singers were accessorized with sequined corsets and significant cleavage.
No significant cleavage of double bonds takes place.
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