Plasmid Constructs for PERK and CHOP

Expression plasmids for human PERK and its DN mutant K621M were obtained from Ronald Wek [48 (link)]. Reporter plasmid pCHOP-Luc, in which luciferase expression is driven by human CHOP promoter (-644 to +91), was provided by Nai Sum Wong [23 (link)]. Reporter plasmids pGRP78-Luc and pGRP94-Luc were gifts from Kazutoshi Mori [40 (link),58 (link)]. The Grp78 and Grp94 promoters are derived from -304 to +34 of human Grp78 gene and -363 to +34 of human Grp94 gene, respectively. Both promoters harbor multiple copies of ER stress response element [58 (link)]. pUPRE-Luc reporter plasmid has been described elsewhere [41 (link),42 (link)].
Mouse monoclonal anti-V5 antibody was purchased from Invitrogen. Mouse anti-FLAG antibody (clone M2) was from Sigma-Aldrich.

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Siu K.L., Chan C.P., Kok K.H., Woo P.C, & Jin D.Y. (2014). Comparative analysis of the activation of unfolded protein response by spike proteins of severe acute respiratory syndrome coronavirus and human coronavirus HKU1. Cell & Bioscience, 4, 3.

Publication 2014

Mouse monoclonal anti-V5 antibody Mouse anti-FLAG antibody (clone M2)

Anti antibody Chop Clone Gifts Grp78 Grp94 Human Luciferase Monoclonal antibody Mouse Plasmid Response element

Corresponding Organization :

Other organizations : University of Hong Kong

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Variable analysis

independent variables

Human PERK expression plasmid
Human PERK DN mutant K621M expression plasmid

dependent variables

Luciferase expression driven by the human CHOP promoter (-644 to +91)
Luciferase expression driven by the human GRP78 promoter (-304 to +34)
Luciferase expression driven by the human GRP94 promoter (-363 to +34)
Luciferase expression driven by the UPRE reporter plasmid

control variables

Antibodies: Mouse monoclonal anti-V5 antibody, Mouse anti-FLAG antibody (clone M2)

controls

None specified
None specified

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