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. Author manuscript; available in PMC: 2011 May 4.
Published in final edited form as: Nat Genet. 2010 Mar 14;42(4):332–337. doi: 10.1038/ng.549

Figure 1.

Figure 1

(a) Cos-7 cells were transfected with pCMV-ORMDL3-GFP. After 24h, cells were fixed using 4% paraformaldehyde and subjected to immunostaining using anti-PDI antibodies as an ER marker. Fluorescence signals were acquired with a confocal microscope. Displayed images represent a single confocal section.

(b) HEK293T cells were co-transfected with a firefly luciferase reporter p5xUPRE-GL3, renilla luciferase as transfection control and pCMV-3XHA or pCMV-ORMDL3-3XHA. After 24h, cells were left untreated or treated using 5μg/ml of tunicamycin or 10μM of thapsigargin for 6h. ORMDL3-3XHA was detected using an anti-HA antibody. Luciferase activities were measured to detect UPRE transcription activation.

(c). HEK293T cells were co-transfected with plasmids encoding shRNA scramble or directed against ORMDL3, firefly luciferase reporter p5xUPRE-GL3 and renilla luciferase as transfection control. After 48h, cells were left untreated or treated using 5μg/ml of tunicamycin or 10μM of thasigargin. ORMDL3 expression was determined by quantitative RT-PCR (left). Luciferase activities were measured to detect UPRE transcription activation (right)