β-hydroxybutyrate modulates neuroinflammatory responses and astrocyte reactivity in an in vitro model of traumatic brain injury

Numerical data obtained from image analysis (immunocytochemistry), Western Blots and antibody arrays measurements.The experiments were performed on primary neuronal-glial cultures, obtained from the cortices and hippocampi of P6-9 Wistar rat pups. Between 14 and 21 DIV, the cells were supplemented with 5 mM DL-β-hydroxybutyrate. On DIV21 the scratch injury was performed with pipette tip. 24h post injury the cultures were fixed for immmunocytochemistry (astrocyte staining - GFAP+DAPI or neurons staining - βIII-tubulin+ DAPI). The density and morphology of neurons and astrocytes in the scratch area was assesed in FIJI. 6h or 24h post-injury the protein was collected for Western Blot analysis of GFAP and NeuN expression. 6h post injury the protein was collected for cytokines experssion analysis, using Rat Neuro Discovery Array C1 (RayBiotech).