Calcium imaging assays
were performed in 96-well plates using the Fluo-4 Direct assay kit
(Invitrogen) and an EVOS FL auto microscope (whole particles) or BMG
Labtech NOVOStar plate reader (particle extracts). Treatment-induced
changes in cellular fluorescence were quantified using the average
value for change in fluorescence, normalized to the maximum response
elicited by ionomycin (10 μM).13 (link),19 (link) In some instances,
the data were also normalized to the prototypical TRPA1 agonist AITC.
Data were also corrected for nonspecific responses, if any, observed
with HEK-293 cells. All agonist, particle, particle extract treatment
solutions were prepared in LHC-9 containing 2% DMSO at a 3× concentration
and added to cells at 37 °C. The final particle concentration
indicated in the figures represents the concentration of the suspension
(or extract) in the treatment well.
were performed in 96-well plates using the Fluo-4 Direct assay kit
(Invitrogen) and an EVOS FL auto microscope (whole particles) or BMG
Labtech NOVOStar plate reader (particle extracts). Treatment-induced
changes in cellular fluorescence were quantified using the average
value for change in fluorescence, normalized to the maximum response
elicited by ionomycin (10 μM).13 (link),19 (link) In some instances,
the data were also normalized to the prototypical TRPA1 agonist AITC.
Data were also corrected for nonspecific responses, if any, observed
with HEK-293 cells. All agonist, particle, particle extract treatment
solutions were prepared in LHC-9 containing 2% DMSO at a 3× concentration
and added to cells at 37 °C. The final particle concentration
indicated in the figures represents the concentration of the suspension
(or extract) in the treatment well.