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. 2018 May 22;9(6):604. doi: 10.1038/s41419-018-0642-6

Fig. 6. Proteasome inhibitors aggravated while protein synthesis suppression ameliorated the UPR and cell death by PPI.

Fig. 6

a, b AGS cells were pretreated with PPI (100 μg/ml) for 24 h in pH 7.4 condition, followed by combination with or without 50 nM Bortezomib or 0.1 μM MG132 for another 24 h. Western blot analysis of apoptosis related protein cleaved-PARP levels (a) and poly-ubiquitinated proteins (b). c, d AGS and HeLa cells were pretreated with CHX (250 ng/ml) for 2 h, and then incubated with PPI (100 μg/ml or 120 μg/ml) for 48 h in pH 7.4 condition. Levels of apoptosis related protein cleaved-PARP, UPR marker CHOP (c), and poly-ubiquitinated proteins (d) were analyzed. e AGS and HeLa cells were pretreated with torin 1 (500 nM) for 2 h, and then incubated with PPI (100 μg/ml or 120 μg/ml) for 48 h in pH 7.4 condition. Indicated proteins were analyzed by western blot. f AGS cells were treated as described in (a). The cell viability was determined by CCK8 assay (left) (**p < 0.01, identified by two-way ANOVA with Sidak’s multiple comparisons test). qRT-PCR analysis of UPR genes was performed (right) (**p < 0.01, for each gene using one-way ANOVA with Tukey’s multiple comparisons test). g AGS cells were treated as described in (c). The cell viability was determined by CCK8 assay (left) (**p < 0.01, identified by two-way ANOVA with Sidak’s multiple comparisons test). qRT-PCR analysis of UPR genes was performed (right) (**p < 0.01, for each gene using one-way ANOVA with Tukey’s multiple comparisons test). h AGS cells were treated as described in (e). The cell viability was determined by CCK8 assay (left) (**p < 0.01, identified by two-way ANOVA with Sidak’s multiple comparisons test). qRT-PCR analysis of UPR genes was performed (right) (**p < 0.01, for each gene using one-way ANOVA with Tukey’s multiple comparisons test)