Measuring Cell ATP Levels

Cell ATP levels were measured as described^{11 (link)}. Assay plates were removed from the incubator and allowed to reach room temperature. CellTiter-Glo (CTG) reagent (Promega) was added (20 µl well⁻¹), with gentle agitation for 30 min. Luminescence as a measurement of cellular ATP levels was read on a PHERAstar Plus microplate reader (BMG Labtech). Results were normalised to those of untreated control cells (no death signal, no MAP4K4 inhibitor) and to 100% cell death (addition of 0.1% Triton X-100, 2 h before CTG). Normalised values were plotted against the log concentration of the death inducer or inhibitor.

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Golforoush P.A., Narasimhan P., Chaves-Guerrero P.P., Lawrence E., Newton G., Yan R., Harding S.E., Perrior T., Chapman K.L, & Schneider M.D. (2020). Selective protection of human cardiomyocytes from anthracycline cardiotoxicity by small molecule inhibitors of MAP4K4. Scientific Reports, 10, 12060.

Publication 2020

CellTiter-Glo (CTG) reagent PHERAstar Plus microplate reader

Assay Cell death Cells Luminescence measurement Map4k4 Promega Triton x 100

Corresponding Organization :

Other organizations : Imperial College London, Domainex (United Kingdom)

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Variable analysis

independent variables

Death inducer or inhibitor concentration (log scale)

dependent variables

Cellular ATP levels (normalized to untreated control and 100% cell death)

control variables

Incubator temperature (room temperature maintained during the assay)
CTG reagent addition (20 µl per well)
Agitation time (30 min)
Luminescence measurement (on PHERAstar Plus microplate reader)

positive control

100% cell death (addition of 0.1% Triton X-100, 2 h before CTG)

negative control

Untreated control cells (no death signal, no MAP4K4 inhibitor)

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