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Fig. 5 | Experimental Hematology & Oncology

Fig. 5

From: Analysis of the anti-proliferative and the pro-apoptotic efficacy of Syk inhibition in multiple myeloma

Fig. 5

a, b Caspase-3 activity is increased significantly upon treatment with Syk inhibitors. a Caspase-3 activity was determined from whole cell lysates by the cleavage of the fluorogenic caspase substrate N-acetyl-Asp-Glu-Val-Asp-aminomethylcoumarin (DEVD-AMC). The release of aminomethylcoumarin was determined by fluorometry using an excitation length of 360 nm and an emission wave-length of 460 nm. Syk-Inhibitors induce a significant concentration-dependent increase of caspase-3 activity. Results are expressed as the mean of three panels per pattern. Results are from one experiment representative of three. DMSO was used as control. The significance is related to DMSO. +P<1.000, *P < 0.1, **P < 0.01, ***P < 0.001. b Western Blot analyses of AMO-1 whole cell lysates treated with Piceatannol was utilized to analyze the expression of pro-caspase 3 and PARP-1 cleavage. After 24 h of incubation with Syk inhibitors, whole cell lysates were prepared and Western Blot analysis was performed. DMSO was used as a control. GAPDH was used to confirm equal amounts of protein. Results are representative of at least three independent experiments. Proteins detected are indicated by an arrow on the right. c Expression of xIAP, Mcl-1, cytochrome c and Survivin upon treatment with Syk inhibitors. Protein extracts were analyzed for the expression of proteins participating in apoptosis such as xIAP, Mcl-1, survivin and cytochrome c by immunoblotting. An increase of cytochrome c was observed, implicating that Piceatannol induces apoptosis via the internal, mitochondrial pathway. There was no effect of Piceatannol on the anti-apoptotic proteins survivin, Mcl-1 and xIAP. Results are representative of various independent experiments. Proteins detected are indicated by an arrow on the right.

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