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cDNA equal to 20 ng of total RNA was amplified by qPCR in duplicate in a ViiA7 qPCR system (Thermo Fisher Scientific), using the Fast Advance Master Mix (Thermo Fisher Scientific), assembled with primer and probe sets for VEGF-A and GFAP genes (IDs: Mm0043736_m1 and Mm01253033_m1 respectively) and one housekeeping gene, 18s rRNA (ID: 4333760 F) (Thermo Fisher Scientific)

In one study, pressure inside a spinal disc dropped to below 100 mm Hg when VAX-D was applied, showing that the treatment can effectively decompress the discs (Ramos et al., 1994)

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