Supplemental Figures

This article contains the following supporting material:

• Supplemental Figure 1.pdf - Expression of atf-6 and pek-1 during development.
  The mRNA levels of atf-6 and pek-1 were measured by RT-PCR using primers shown in the diagram. Cloned competitor DNA was generated by using a genomic fragment that could be discerned from the RT-PCR cDNA product on an agarose gel. The indicated amount of cloned competitor was added to either total RNA (top gel) or polyA(+) mRNA (bottom gel) from L1 larvae or adult nematodes. The amplification products were run on an agarose gel as shown. The gel on the left shows products for atf-6, whereas the gel on the right shows products for pek-1.
  
  
• Supplemental Figure 2.pdf - Expression of xbp-1 from the XB3 and XB7 transgenes.
  The mRNA level and product size of xbp-1 was measured by RT-PCR from either non-transgenic nematodes, nematodes carrying the XB3 transgene, or nematodes carrying the XB7 transgene using primers shown in the diagram. The amplification products were run on an agarose gel as shown. Lanes 1-3 used primers that should only amplify XBP-1B mRNA from the endogenous gene. Lanes 4-6 used primers that should amplify XBP-1A or XBP-1B mRNA, but only from the XB3 and XB7 transgenes. The size of molecular weight markers is indicated.
  
  
• Supplemental Figure 3.pdf - Splicing of the regulatory intron in the absence of ER stress.
  The mRNA product sizes of xbp-1 were measured by RT-PCR from either nematodes carrying the XB3 transgene or nematodes carrying the XB7 transgene. Products from either XB3 or XB7 RNA from Supplemental Figure 2 were subcloned and used as template in a second PCR reaction using oligos R74.3-5 and R74.3-4 to measure the size of the regulatory intron (indicated in red). Unspliced products yield a 160 bp band, whereas spliced products yield a 147 bp product. The amplification products were run on an agarose gel as shown. *Lanes with the 147 bp product, indicating splicing.
  
  
• Supplemental Figure 4.pdf - Sequence of xbp-1 genomic DNA and cDNA.
  The RT-PCR products from Supplemental Figure 2 were sequenced and aligned to the xbp-1 genomic sequence. The gray boxes indicate common introns that are spliced out of both XBP-1A and XBP-1B message. The yellow box indicates the regulatory intron that is spliced out by IRE-1.
  

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