10.1371/journal.pone.0031944.g004
Adrian A. Moreno
Adrian
A. Moreno
M. Shahid Mukhtar
M.
Shahid Mukhtar
Francisca Blanco
Francisca
Blanco
Jon Lucas Boatwright
Jon
Lucas Boatwright
Ignacio Moreno
Ignacio
Moreno
Melissa R. Jordan
Melissa
R. Jordan
Yani Chen
Yani
Chen
Federica Brandizzi
Federica
Brandizzi
Xinnian Dong
Xinnian
Dong
Ariel Orellana
Ariel
Orellana
Karolina M. Pajerowska-Mukhtar
Karolina
M. Pajerowska-Mukhtar
bZIP60 mRNA splicing is stimulated by chemicals that trigger the UPR.
Public Library of Science
2012
mrna
splicing
stimulated
chemicals
2012-02-16 00:47:16
Figure
https://plos.figshare.com/articles/figure/_bZIP60_mRNA_splicing_is_stimulated_by_chemicals_that_trigger_the_UPR_/352836
<p><b>A,</b> Schematic representation of two approaches used to detect the bZIP60 mRNA spliced forms. Primers sets flanking the putative splicing regions (solid arrows) are indicated (Top) to amplify bZIP60u and bZIP60s forms using RT-PCR. Alternative, RT-PCR products are subsequently digested using Alw21I restriction enzyme (Bottom). The latter approach will highlight the length differences between bZIP60u and bZIP60s since the Alw21I restriction site is present in bZIP60u and absent in bZIP60s. bZIP60u and bZIP60s PCR products upon digestion are shown. <b>B,</b> Processing of bZIP60 mRNA was analyzed by gel electrophoresis in agarose (3.5% p/v). RT-PCR products (Top) or RT-PCR products digested with Alw21I (bottom) were obtained from RNA samples of Arabidopsis seedlings (6-day-old) treated for 2 hours with several chemicals that trigger the UPR (Tm 5 µg/mL; DTT 5 mM; CPA 100 µg/mL; Thapsigargin 500 nM). DMSO and water-treated samples served as mock controls for chemicals. Asterisk indicates a hybrid band formed by the bZIP60u and bZIP60s PCR products. Such hybrid band has been also observed and documented in RT-PCR analysis of XBP-1 processing <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031944#pone.0031944-Shang1" target="_blank">[56]</a>. Elongation factor 1 alpha (EF1α) expression served as a control. All the experiments were performed at least three times with similar results.</p>