Abstract

<p>(A) mRNA-Seq expression values of 24/141 mRNAs identified (<a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1002277#pbio.1002277.g003" target="_blank">Fig 3B–3D</a>) to be both <i>Ire1α-</i> and high glucose-dependent for their induction ([<i>n</i> = 5], [<i>p</i>-values ≤ 0.01]). (B) Autoradiograph from whole cell islet lysates prepared by steady-state (18 h) [<sup>35</sup>S]-Cys/Met radiolabeling from 12 wk post-Tam mice (<i>n</i> = 3) following peptide gel electrophoresis. (C) Western blotting for proinsulin/preproinsulin, IRE1α, SEC11C, SSR1, and tubulin after peptide gel electrophoresis of lysates prepared 72 h after COS-1 cells were coinfected with adenoviruses expressing <i>WT</i> preproinsulin, the Akita mutant (A), and/or the (G) GFP, (X) XBP1s, or (Δ) the dominant negative IRE1α-RNase mutant K907A representative results shown (<i>n</i> = 4). (D) Ribosomes and their relative position to one another were measured from electron micrographs at a magnification of 25,000x. Total numbers of ribosomes analyzed are shown below the images ([<i>n</i> = 3], [<i>p</i> = 2.2 x 10<sup>−15</sup>]). (E) Subcellular fractionation and western blot analysis for ribosomal small subunit 9 and tubulin of the <i>Ire1α</i><sup><i>Fe/Fe</i></sup> β cell insulinoma line at after 2-h glucose shift from 12 mM to either 4 mM or 36 mM with or without infection by the indicated adenoviruses were blotted representative of (<i>n</i> = 3). The 12 mM condition western blots and the quantified results normalized to tubulin membranous/cytosolic are shown in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1002277#pbio.1002277.s010" target="_blank">S6D Fig</a>.</p

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