HDAC2 inhibition reversed the downregulated expression of NR2B caused by maternal isoflurane exposure.

(A1) Amplification plot of NR2B and β-actin mRNA; (A2) Agarose gel electrophoresis images of NR2B and β-actin mRNA; (A3) Maternal isoflurane exposure downregulated the expression of NR2B mRNA (mean ± SE): The levels of NR2B mRNA in isoflurane exposure group (I2N, I4N, I8N) were lower than CN group (* p < 0.05). The NR2B mRNA levels in I4N and I8N group were lower than I2N group (# p < 0.05); (A4) SAHA reversed the downregulated expression of NR2B mRNA (mean ± SE): The levels of NR2B mRNA in I2S, I4S and I8S subgroup were higher than I2N, I4N and I8N subgroup respectively (* p < 0.05). The NR2B mRNA levels in I8S subgroup were lower than CN group (# p < 0.05). (B1) NR2B protein western blot images; (B2) Maternal isoflurane exposure downregulated the expression of NR2B protein (mean ± SD): The protein levels of NR2B protein were lower than CN group (# compared with I2N group, p<0.05b); (B3) HDAC2 inhibition reversed the downregulated expression of NR2B protein: The protein levels of NR2B in I2S, I4S and I8S subgroups were higher than I2N, I4N and I8N sugroups respectively. The levels of NR2B protein in I8S subgroup were lower than CN group (# p < 0.001). This indicates that SAHA cannot completely reverse the downregulated effect caused by isoflurane when the exposure time prolonged to 8 hours.