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Fig. 5 | BMC Biology

Fig. 5

From: Optimized photo-stimulation of halorhodopsin for long-term neuronal inhibition

Fig. 5

Mechanistic insight of the inactivation and recovery of eNpHR3.0. a Decreasing extracellular proton concentration enhances inactivation of eNpHR3.0. Currents were measured in the same oocyte in Ringer’s solution at pH 5.6, pH 7.6, or pH 9.6. b Increasing extracellular chloride concentration reduces inactivation of eNpHR3.0. Currents were measured in the same oocyte at different chloride concentrations. Buffers with different chloride concentrations were achieved by mixing Ringer’s solution (pH 7.6) and NMG-Asp solution (pH 7.6) at different ratio. c Recovery of eNpHR3.0-mediated photo-currents in Ringer’s solution at pH 5.6 (n = 4 cells), pH 7.6 (n = 8 cells), or pH 9.6 (n = 4 cells) at a holding potential of − 40 mV. d Recovery of eNpHR3.0-mediated photo-currents at an extracellular chloride concentration of 6 mM (n = 5 cells), 16 mM (n = 6 cells), 60 mM (n = 6 cells), or 121 mM (n = 5 cells). pH was set to 7.6 and holding potential to − 40 mV. Dotted lines in c and d represent bi-exponential fits to population data. e Recovery of eNpHR3.0 (pH 7.6) at holding potentials of − 100 mV (n = 7 cells), − 40 mV (n = 5 cells), or + 20 mV (n = 5 cells). Five hundred ninety-nanometer light at an intensity of 2.6 mW/mm2 was applied for 60 s in a and b, while in ce, additional 10-ms 590-nm light pluses at the same intensity were delivered at 1, 5, 10, 20, 40, 60, 120, and 300 s after the initial 60-s illumination, as in Fig. 4e. Data are presented as mean ± SEM. *P < 0.05, ***P < 0.001. a, b Asterisks indicate significance levels of post hoc t tests with Bonferroni correction following one-way repeated-measures ANOVA

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