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Diluted siRNA was mixed with transfection reagent and transfected into cells seeded in 24-well plates

Safety of every-other-day fasting in the treatment of spinal cord injury: a randomized controlled trial

The final pellets were suspended in Laemmli sample buffer, boiled for 5 min, and then briefly centrifuged to collect the supernatants, which were then subjected to SDS-PAGE followed by immunoblotting analysis