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Western blot analysis involved preparing whole cell lysates using RIPA lysis buffer, quantifying protein concentration via BCA assay, followed by SDS-PAGE, transfer to nitrocellulose membranes, incubation with primary and HRP-conjugated secondary antibodies, and detection using an ECL solution [1]

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While the VPA-induced ASD model effectively recapitulates neurodevelopmental and gastrointestinal phenotypes associated with prenatal environmental insults [43], we acknowledge it does not fully represent the genetic heterogeneity of human ASD