Xbp1 Expression Quantification Protocol

As described previously (Rutkevich et al., 2010 (link)), total RNA was isolated from cells using the RNeasy kit (Qiagen) and subjected to one-step reverse transcription PCR (Qiagen) using either human or mouse Xbp1 amplification primers: human, 5′-GGAGTTAAGACAGCGCTTGG-3′ and 5′-GAGATGTTCTGGAGGGGTGA-3′; mouse, 5′-GGCCTTGTGGTTGAGAACCAGGAG-3′ and 5′-GAGTCTGATATCCTTTTGGGCATTC-3′. As a positive control, cells were incubated with 10 μg/ml tunicamycin overnight.

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Stocki P., Sawicki M., Mays C.E., Hong S.J., Chapman D.C., Westaway D, & Williams D.B. (2016). Inhibition of the FKBP family of peptidyl prolyl isomerases induces abortive translocation and degradation of the cellular prion protein. Molecular Biology of the Cell, 27(5), 757-767.

Publication 2016

RNeasy kit one-step reverse transcription PCR

Cells Human Mouse Primers Reverse transcription Tunicamycin Xbp1

Corresponding Organization : University of Toronto

Other organizations : University of Alberta

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Variable analysis

independent variables

Incubation with 10 μg/ml tunicamycin overnight

dependent variables

Expression of human or mouse Xbp1

control variables

Cells not incubated with tunicamycin

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