Fig. 6

IRE1 and XBP1 are partially required for Tg-induced caspase activation and upregulation of DR5-, but not LC3B expression in LNCaP cells. a LNCaP cells were transfected for 2 d with the indicated siRNAs (siCtrl = non-targeting control siRNA), employing two different siRNA oligoes for each target (designated by − 1 and − 2). After 30 h of treatment with 0.02% DMSO (“DMSO”, also transfected with siCtrl) or 100 nM Tg in the absence of presence of 0.5 μM JNK inhibitor JNK-IN-8 (JNKi, also transfected with siCtrl), whole cell lysates were prepared and subjected to western blotting with the indicated antibodies; Casp3 = caspase-3 (only cleaved caspase-3 bands are shown), cl-PARP = cleaved PARP, p-JNK = phospho-JNK. The positions of molecular weight markers are indicated to the left. In the presence of JNK-IN-8, the two p-JNK bands shift towards slightly slower migration due to covalent binding of the inhibitor to the JNK1 and JNK2 isoforms. One representative blot out of at least 3 independent experiments. b-g Quantifications of western blots from (a), normalized to the tubulin loading control and then to the siCtrl+Tg condition. Mean ± SEM from 4 (b-f) or 3 (g) independent experiments. Dots represent individual values, with a separate color for each experiment. *P < 0.05, **P < 0.01, ***P < 0.001, ns; not significant, One-way ANOVA compared to the Tg + siCtrl condition