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. 2013 Jul 14;4(8):1212–1229. doi: 10.18632/oncotarget.1130

Figure 2. Effect of SMIP004 on the unfolded protein response (UPR).

Figure 2

(A) Lysate from LNCaP-S14 cells treated with increasing concentrations of SMIP004 for 24 h were analyzed for UPR markers by immunoblotting. (B) Kinetics of UPR induction by 40 μM SMIP004. (C) XBP1 splicing was analyzed by RT-PCR with total RNA from LNCaP-S14 cells treated with increasing concentrations of SMIP004 for 24 h. (D) Kinetics of XBP1 splicing in response to 40 μM SMIP004. (E) Cells were treated with increasing concentrations of SMIP004 for 24 h in the presence of increasing concentrations of SP600125. JNK phosphorylation and PARP cleavage were analyzed by immunoblotting. (F) Cells were treated with increasing concentrations of SMIP004 for 24 h in the presence of increasing concentrations of SB203580. p38 phosphorylation and PARP cleavage were analyzed by immunoblotting. (G) DNA fragmentation was measured in extracts from cells treated with SP600125 (40 μM) or SB203580 (40 μM) and SMIP004 (40 μM) for 24 h. The graph represents the DNA fragmentation enrichment factor ± standard deviations (normalized to DMSO treatment) of three replicates. (H) Proliferation of cells transfected with specific siRNA for IRE1 and CHOP and treated with SMIP004 (10 μM) for 48 h was measured by MTT assay. The graph represents the mean ± standard deviations of four replicates.