Fig. 7

IRE1 does not oppose Tg-induced upregulation of DR5 mRNA. a-d LNCaP (a and b) or HCT116 (c and d) 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). Subsequently, cells were treated with 100 nM Tg or 0.01% DMSO (also transfected with siCtrl) for 6 h, and subjected to real-time RT-PCR to quantify DR5 mRNA levels (a and c) or western blotting to validate efficient knockdown of IRE1 and XBP1s protein levels (b and d). In (a and c), relative mRNA levels are shown normalized to the DMSO control-treated condition (set to 1 and indicated by the dotted line in the graphs), i.e. the conditions shown are all with Tg treatment. Mean ± SD of triplicate measurements from one representative experiment out of two. Of note, the reduction observed with siXBP1–1 in (a) was not reproducible. In (b and d), one representative blot out of 2 independent experiments is shown