Monitoring Hsf1 Activity via yNlucPEST

A yNlucPEST reporter was used to monitor Hsf1 activity as previously described^{49 (link)}. Briefly, Nano-Glo substrate (Promega, Madison, WI, USA) was diluted 1:100 with the supplied lysis buffer and mixed 1:10 with logarithmic growing cells carrying the reporter plasmid pCA955. Bioluminescence was determined after 3 min of incubation, using an Orion II Microplate Luminometer (Berthold Technologies GmbH, Bad Wildbad, Germany) and the obtained signal was normalized to OD₆₀₀.

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Minoia M., Quintana-Cordero J., Jetzinger K., Kotan I.E., Turnbull K.J., Ciccarelli M., Masser A.E., Liebers D., Gouarin E., Czech M., Hauryliuk V., Bukau B., Kramer G, & Andréasson C. (2024). Chp1 is a dedicated chaperone at the ribosome that safeguards eEF1A biogenesis. Nature Communications, 15, 1382.

Publication 2024

Nano-Glo substrate Orion II Microplate Luminometer

Corresponding Organization :

Other organizations : Stockholm University, DKFZ-ZMBH Alliance, Heidelberg University, Rigshospitalet, Science for Life Laboratory, Lund University

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Variable analysis

independent variables

Nano-Glo substrate dilution (1:100 with supplied lysis buffer)

dependent variables

Bioluminescence

control variables

Mixing ratio of Nano-Glo substrate and cells (1:10)
Incubation time (3 minutes)
Cell growth phase (logarithmic growth)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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