METHODS AND ETHICAL COMPLIANCE
An 8 year-old girl diagnosed with DEE underwent trio whole-exome sequencing. HEK293T and HeLa cells were transfected with CHRM1 wild-type and mutated plasmids tagged at C-terminal. Western blot and immunoblotting assays were done to study protein expression and cell trafficking. Luciferase reporter assays were performed to measure cAMP and calcium cell signaling activity. GraphPad software was used to perform the statistic analysis (See Supplementary Information). The study was approved by the local Ethics Committee (PR(AG)223/2017) and informed consent was obtained from the patient’s parents according to the Helsinki declaration.