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Intracellular Ca 2+ imaging Cells were transfected using Nucleofector as described before using the plasmids pCMV_R-GECO1 (Addgene #32444, measures intra-cytosolic Ca 2+ , red fluorescence) and pCMV_CEPIA2mt (Addgene #58218, measures intra-mitochondrial Ca 2+ , green fluorescence)

As directed by a healthcare professional

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Such abnormalities have been reported in the ASD population previously 52, 53, 54 and zinc supplementation (along with B6) has been shown to decrease copper levels and improve function in ASD in an uncontrolled study