Abstract

<p>(A,B) ER-stress induction of both Siah1 and Siah2 mRNA is attenuated in <i>Atf4<sup>−/−</sup></i> MEFs (A) and <i>Ire1α<sup>−/−</sup></i> MEFs (B). Littermate-matched MEFs of the indicated genotypes were subjected to treatment with TM (2 µg/ml) or TG (1 µM) for 6 h and the relative expression of Siah1 and Siah2 mRNA was measured by qPCR. (C) IRE1α is required for ER-induced Siah2 mRNA levels. Ectopic expression of sXBP1 restores TM-induction of Siah2 mRNA in <i>Ire1α<sup>−/−</sup></i> MEFs. <i>Ire1α<sup>−/−</sup></i> MEFs were infected either with adenovirus encoding either β-gal or sXBP1. After 24 h, RNA was prepared and quantified using qPCR for the relative levels of Siah2 mRNA. (D) ER-stress induction of Siah2 mRNA levels but not of Siah1 mRNA are attenuated in <i>Atf6α<sup>−/−</sup></i> MEFs. Littermate-matched MEFs of the indicated genotypes were subjected to treatment with TM (2 µg/ml) TG (1 µM) for 6 h and the relative expression of Siah1 and Siah2 mRNAs were measured by qPCR. (E) ER Stress induction of Siah1/2 transcripts is not HIF1-dependent. WT and HIF1α KO MEFs were subjected to TM (2 µg/ml) or TG (1 µM) treatment and RNA prepared 6 h later was subjected to analysis of Siah1 or Siah2 transcripts. *** p<0.0005, ** p<0.005, * p<0.07 compared to Ire1 KO (C) or to WT under the same condition (student's t test). The results are shown as the mean values ± S.E. of three independent experiments.</p

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