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Western blot analysis RIPA Lysis Buffer (TransGen, Beijing, China) was used to extract total protein from tissues/fibroblasts and a Nuclear and Cytoplasmic Protein Extraction Kit (TransGen) was used to extract the protein from nucleus and cytoplasm of fibroblasts

Module gene analysis and GO term mainly pointed to a role of RAMP1 and RAMP3 in impacting neuronal and synaptic function but as these receptors are widely expressed, their role is not limited to a specific type of neurons

Monitor for any return of symptoms as a signal to reduce intensity

Samples are then rinsed inPBST2 times for 10 minutes each then incubated in alexa-488 conjugated secondary antibodies for 2 hours at room temperature or overnight at 4 C