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Cell were washed and placed in normal Ringers buffer (160 mM NaCl, 4.5 mM KCl, 2 mM CaCl 2 , 1 mM MgCl 2 , 10 mM HEPES, pH 7.4) or Low Ca 2+ Ringers where CaCl 2 was omitted and 1 mM Ethylenediaminetetraacetic acid (EDTA) added

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A library for RNA-seq was prepared using the Ovation Human FFPE RNA-Seq Library Systems (NuGEN) (for whole worm samples) and the QuantSeq 3 mRNA-Seq Library Prep Kit for Illumina (FWD) (Lexogen) (for intestinal samples)

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It is possible that these variants have impaired folding and localisation, although R169W has previously been shown to localise normally to the membrane 53

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