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. 2015 Mar 17;16(3):420–429. doi: 10.1080/15384047.2014.1003003

Figure 4.

Figure 4

(See previous page). MLN4924-induced apoptosis is mediated through the intrinsic apoptotic signaling and associated with a coordinated dysregulation of pro-apoptotic and anti-apoptotic proteins. (A) MLN4924 induced apoptosis at early time post treatment. SU-DHL-4 and Toledo cells were treated with MLN4924 at 0.1 and 0.3 μM or DMSO for indicated time points, and subjected to immunoblotting using the antibody against cleaved PARP with GAPDH as a loading control. (B) MLN4924 induced cell apoptosis mainly via the intrinsic apoptotic signaling pathway. SU-DHL-4 and Toledo cells, treated with MLN4924 at 0.1 and 0.3 μM or DMSO for 48 h, were subjected to immunoblotting using antibodies against caspase-8 and cleaved caspase-9 with GAPDH as a loading control. (C) Effects of MLN4924 on expression of pro-apoptotic and anti-apoptotic proteins. SU-DHL-4 and Toledo cells were treated with MLN4924 at 0.1 and 0.3 μM or DMSO for 48 h, followed by immunoblotting using indicated antibodies against pro-apoptotic (left panel) and anti-apoptotic (right panel) proteins with GAPDH as a loading control. (D) Effects of MLN4924 on transcriptional activation of apoptosis-regulatory proteins. Toledo cells, treated with 0.3 μM MLN4924 or DMSO for 36 h, were subjected to the real-time PCR analysis using human apoptosis PCR array as described in Materials and Methods (n = 3).