Calcium flux/TRPV3 activity
was assayed using the Fluo-4 Direct assay kit (Invitrogen). Treatment-induced
changes in cellular fluorescence were quantified from fluorescence
micrographs or using a NOVOStar fluorescence plate reader (BMG Labtech;
Offenberg, Germany), as previously described.30 (link)−34 (link) All agonist treatment solutions were prepared in
LHC-9 at 3× concentration and added to cells at 37 °C. Particles
were added based on the surface area of the vessel (μg/cm2). Data from HEK-293 TRP-over-expressing cells were corrected
for non-specific responses, if any, observed with HEK-293 cells. Data
were also either normalized to the maximum attainable change in fluorescence
elicited by ionomycin (10 μM) post-treatment or to the change
elicited by a maximum stimulatory concentration of a positive control
agonist, as noted in the figure legends.
was assayed using the Fluo-4 Direct assay kit (Invitrogen). Treatment-induced
changes in cellular fluorescence were quantified from fluorescence
micrographs or using a NOVOStar fluorescence plate reader (BMG Labtech;
Offenberg, Germany), as previously described.30 (link)−34 (link) All agonist treatment solutions were prepared in
LHC-9 at 3× concentration and added to cells at 37 °C. Particles
were added based on the surface area of the vessel (μg/cm2). Data from HEK-293 TRP-over-expressing cells were corrected
for non-specific responses, if any, observed with HEK-293 cells. Data
were also either normalized to the maximum attainable change in fluorescence
elicited by ionomycin (10 μM) post-treatment or to the change
elicited by a maximum stimulatory concentration of a positive control
agonist, as noted in the figure legends.
