Nuclear and cytoplasmic protein extractions of E17 neurons were carried out using an NE-PER kit (catalog no. 78833, Thermo Fisher Scientific) as per the manufacturer’s instructions, with an extra wash with cytoplasmic extraction reagent (CER1) before addition of buffer nuclear extraction reagent (NER1). Four hundred micrograms of protein was used per sample. CysNO treatment for 20 min was carried out in the CER1 (cytoplasmic) or NER1 (nuclear) buffer with intermittent mixing. CysNO was removed after treatment either by acetone precipitation or using P6 spin columns prewashed in HEN buffer. Protein amounts were equalized by bicinchoninic acid (BCA) assay, and then, the volume was made up to 500 μl with HEN buffer. SNORAC was carried out according to (15) with the following modifications. Thiopropyl sepharose 6B resin (catalog no. 17-0420-01, GE Healthcare) was used, after three washes in BPC water and one wash in HEN buffer. Beads were centrifuged at 500g for 10 min at 4°C in between washes. Beads were stored (no longer than 1 day) in HEN buffer at 4°C in a 1:1 ratio. MS samples were frozen with HEN buffer diluted 10-fold in water and stored at −20°C.

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