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. 2018 Sep 3;150(9):1273–1286. doi: 10.1085/jgp.201711961

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© 2018 Zhou et al.

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Figure 5. — Applications of a popularly used photosensitizer, Rose Bengal, and ¹O₂ quenchers (sodium azide and Trolox-C) support the involvement of ¹O₂. (A) Schematic drawing of the experiment configuration. Rose Bengal was applied to the bath solution and in contact with the intracellular side of the channel. (B) In the presence of 100 nM Rose Bengal, laser pulses were applied in the middle of the voltage step from 0 to −100 mV. The current traces recorded before the application of laser pulses are shown below. (C) In the presence of 100 nM Rose Bengal, laser pulses were applied preceding the voltage step to −100 mV. The current traces recorded before the application of laser pulses are shown below. (D) Averaged results showing the effect of laser pulses (during the voltage step) on the I_macro amplitude of the WT (n = 8, black) and spHCN/H462A mutant (n = 8, red) channels. (E) Averaged results showing the effect of laser pulses (before the voltage step) on the I_macro amplitude of the WT (n = 5, black) and spHCN/H462A mutant (n = 5, red) channels. (F) Averaged results showing the effect of sodium azide (n = 7, blue) and Trolox-C (n = 8, purple) on the photodynamic modification of WT spHCN channels. FITC-cAMP was the photosensitizer. (G) Averaged results showing the effect of Trolox-C (purple) on the photodynamic modification (n = 11) of WT spHCN mutant (n = 7, red) channels. Rose Bengal was the photosensitizer. For D–G, results are mean ± SEM; ***, P ≤ 0.001. Error bars represent SEM.