The effects of the antagonists DHβE, derquantel and α-BTX (10 μM) were examined in the presence 100 μM ACh as previously described for Asu-ACR-16 [29]. In short, X. laevis oocytes co-injected with Tsu-acr-16-like- and Xla-ric-3 cRNA were sequentially superfused with ACh for 10 s, then ACh + antagonist for 10 s, and finally with ACh for 10 s. For α-BTX a five-step protocol including a pre-incubation (10 s) with the antagonist (10 μM) was used with ACh (100 μM). Xenopus laevis oocytes, co-injected with Tsu-acr-16-like- and Xla-ric-3 cRNAs, were exposed to: i) a control application of 100 μM ACh for 10 s (first application); ii) followed by a wash-off period of 5 min; iii) then by an application of 10 μM α-BTX for 10 s, immediately followed by 100 μM ACh and the continued presence of α-BTX for 10 s (second application); iv) then a wash-off period of 5 min; v) and finally an application of ACh for 10 s (third application). Control oocytes were exposed to ACh for 10 s in 3 consecutive steps, each separated by a wash-off period of 5 min. For each antagonist, n = 6–8.

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