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Although K+ blockers were added to the application solution, the recording was contaminated by voltage-gated K+ currents (Fig. 1b).
To test this possibility we measured the steady-state inactivation of the current when the R-type blocker, SNX-482 (30 nM) was added to the application solution.
The following toxins and blockers were used: nifedipine (Sigma) was diluted in 95% ethanol immediately before use and the application solution was protected from ambient light.
In experiments with toxins, 0.1 mg/ml bovine serum albumin (BSA, Sigma) was added to the application solution to prevent non-specific binding.
To reduce the contamination by K+ currents and increase the amplitude of the inward current we replaced Ca2+ ions (2 mM) in the application solution with Ba2+ ions (5 mM) (Fig. 1c).
To isolate the Q-type current the blockers for L-, N-, R- and P-type channels were added to the application solution before blocking the Q-type channel (Fig. 5e).
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The application solutions containing various concentrations of choline were prepared fresh daily.
The application solutions with the different toxins and blockers were applied locally using perfusion tubing coated with Sigmacote (Sigma) to prevent binding of the toxins.
A sample of the pesticide application solution was not available for H1; however, based on the chlorpyrifos levels measured in the indoor air postapplication, the applied amount in H1 was probably similar to amounts applied in homes H2 H7.
Spray the inside of the window generously with application solution.
Next, peel off the film liner and spray the exposed adhesive with application solution.
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