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.