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To introduce the GFP-P2A-BFP insert, primers containing 15-bp overlaps with each of the FSP1 homology arms were used to amplify a codon-optimized GFP-P2A-BFP gBlock (Integrated DNA Technologies)

This is not just a theoretical issue

Representative blots of n = 3 shown

Studies focused on traumatic brain injury recovery and aging models have used 48-hour interval protocols effectively, suggesting this represents near the minimum frequency that maintains cumulative neuroplastic stimulus