Figure 1. Suppression of UPR genes by Zhangfei in ONS-76 medulloblastoma cells treated with thapsigargin.

A, B, C – “Volcano” plots from data derived from qRT-PCR arrays designed to monitor transcripts of genes associated with the UPR. The horizontal axes in these graphs represent log₂ fold differences between the samples indicated. The vertical axis represents -log₁₀ of P values for data derived from three pairs of arrays. The horizontal line (-1.3) represents a P value of 0.05. Transcripts that changed more than 2 fold (<-2 or >2, vertical lines) and had a P value <0.05 were considered significant and are indicated by solid spots with gene designations. A. Difference in UPR transcripts between ONS-76 cells expressing either Zhangfei or LacZ. Cells were infected with adenoviruses expressing either Zhangfei or LacZ and harvested 48 hr later for RNA extraction and analysis. B. Differences between LacZ-expressing cells treated with either thapsigargin or DMSO (solvent for thapsigargin). Cells were treated for 4 hr then infected with adenovirus vector expressing LacZ and harvested for RNA extraction and analysis 48 hr later. C. Effect of Zhangfei on UPR genes activated by thapsigargin. Cells were treated with DMSO or thapsigargin for 4 hr then infected with adenovirus vectors expressing either Zhangfei or LacZ and harvested 48 hr later for RNA extraction and analysis. D. A comparison of data from the qRT-PCR arrays (C.) with data from qRT-PCR experiments using primers designed “in-house”. ATF4 – activation transcription factor 4CEBPB - CCAAT enhancer binding protein-beta, DDIT3 – DNA damage inducible transcript -3, DNAJB9 – homologue of DNAJ/ 40 kD heat shock protein, EDEM – ER degradation enhancer mannosidase alpha-like 1, ERN1 – ER to nucleus signaling, HERPUD1 – homocysteine-inducible ER stress inducible ubiquiti-like domin member 1, HSPA1B – heat shock 70 kD protein 1B, INSIG1 – insulin-induced gene 1, MAPK10 – mitogen-activated protein kinase 10, Xbp1 - X box binding protein 1.