Evaluating TRPA1 Activation by cdPM

Activation of TRPA1 by cdPM and cdPM extracts was evaluated by quantifying changes in intracellular calcium content in human TRPA1-overexpressing HEK-293 cells using the Fluo-4 Direct assay kit (Invitrogen, Carlsbad, CA), as previously described [40 (link),41 (link)]. Data are represented as the percentage of the maximum change in cellular fluorescence achieved by ionomycin treatment (10 μM) and normalized to responses elicited by a positive control (allyl-isothiocyanate, 150 μM). To determine TRPA1 involvement in the biological effects of cdPM, lung cells were co-treated with the TRPA1 antagonist HC-030031 (25 μM).

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Jaramillo I.C., Sturrock A., Ghiassi H., Woller D.J., Deering-Rice C.E., Lighty J.S., Paine R., Reilly C, & Kelly K.E. (2017). Effects of Fuel Components and Combustion Particle Physicochemical Properties on Toxicological Responses of Lung Cells. Journal of environmental science and health. Part A, Toxic/hazardous substances & environmental engineering, 53(4), 295-309.

Publication 2017

Fluo-4 Direct assay kit

Allyl isothiocyanate Assay Biological Calcium Cells Fluo 4 Fluorescence Hc 030031 Hek 293 cells Human Intracellular Ionomycin Lung

Corresponding Organization : University of Utah

Other organizations : Boise State University

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Variable analysis

independent variables

CdPM and cdPM extracts

dependent variables

Changes in intracellular calcium content in human TRPA1-overexpressing HEK-293 cells

control variables

Fluo-4 Direct assay kit (Invitrogen, Carlsbad, CA)
Ionomycin treatment (10 μM)
Allyl-isothiocyanate (150 μM)
TRPA1 antagonist HC-030031 (25 μM)

positive control

allyl-isothiocyanate (150 μM)

negative control

TRPA1 antagonist HC-030031 (25 μM)

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