Fig. 5.

Effect of nicotinamide or saline on p65 translocation; on p65, IL-1β and TNF-α mRNA levels, and on ELISA TNF-α protein levels in mesencephalon of asphyxia-exposed and control rats. Caesarean-delivered control (CS) and asphyxia-exposed (AS) rats were treated with a single dose of nicotinamide (0.8 mmol/kg, i.p.) (CNam, ANam) or saline (0.1 ml, i.p.) (CSal, ASal), 1 h after delivery, euthanized at 8 or 24 h after birth. Selected brain tissue was treated for Western blots, RT-qPCR or ELISA. a Representative immunoblots for p65, α-tubulin and histone H4 levels, measured in cytoplasmic (c) and nuclear (n) protein extracts. b Nuclear p65 levels normalized to total p65. c De novo synthesis of p65, d IL-1β and e TNF-α, measured by RT-qPCR 8 h and/or 24 h after delivery (data analysed in triplicates with MxPro software, normalized to GAPDH mRNA levels). f TNF-α protein levels measured in total protein extracts with Quantikine^® ELISA Rat (normalized by mg of protein for each sample, in duplicate) (CSal open columns, CNam dashed columns, ASal grey columns, ANam doubled dashed columns). Pair-wise comparisons analysed with Student t test (*p < 0.05, **p < 0.005)