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. 2007 Jun 27;2(6):e571. doi: 10.1371/journal.pone.0000571

Figure 5. Monitoring of ER stress with Gluc.

Figure 5

293T cells expressing Gluc were subjected to no or different concentrations of DTT (A–D) or thapsigargin (E–F) to induce ER stress. (A) Real-time RT-PCR for spliced XBP-1 mRNA which increased in response to >1 mM DTT 4 hrs after treatment. Fold induction is calculated with respect to the control non-infected/non-treated cells. (B&F) Western blot analysis showing upregulation of BiP and/or phosphorylated eIF2alpha levels in response to ER stress 24 hrs after treatment. Blot is also probed with Gluc antibody as well as beta-tubulin antibody for equal loading. (C&E) Conditioned medium assayed for Gluc activity 4 hrs after DTT or thapsigargin treatment showing that Gluc secretion is decreased in response to ER stress. (D) Untreated or treated cells with 1 mM DTT were monitored overtime for the level of Gluc secretion by assaying an aliquot of the conditioned medium for Gluc bioluminescence. The mean ± S.E.M. is presented on the graphs (n = 3), with *p≤0.01 as calculated by the student's t-Test.