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. 2014 Sep 25;10(9):e1004675. doi: 10.1371/journal.pgen.1004675

Figure 7. Prolonged ER stress increases Derlin-1 proteins that are not bound to TER94.

Figure 7

(A) Confocal micrographs of retinas from GMR>xbp1-eGFP flies fed with different concentrations of Tunicamycin (Tm). The ER stress response to Tm treatment is dose-dependent, with 24 µM Tm eliciting robust Xbp1-eGFP signals. (B) Western analyses of lysates from flies fed with 24 µM Tm for 2, 4, and 6 days. The levels of endogenous Derlin-1 and TER94 (revealed by anti-Derlin-1 and anti-VCP antibodies) in response to continuous Tm treatment are compared to those from untreated (con). The β-Tubulin level is included as loading control. In the bar graph, endogenous Derlin-1 and TER94 levels (as shown in the Western in B) are normalized to loading controls and presented in fold change as compared to untreated control. Values shown represent mean ± SE from five independent experiments. *p<0.05; **p<0.01 (one-way ANOVA with Bonferroni's multiple comparison test). (C) Sequential IP (1st IP by anti-VCP and 2nd IP by anti-Derlin-1) of lysates from flies fed with 24 µM Tm for 2, 4, and 6 days. The immunoprecipitates from control and Tm-treated flies are probed with anti-VCP and anti-Derlin-1.