Figure 6.

Melatonin Increases PIAS1 Expression and AICD SUMOylation and Increases the Expression of NEP and TTR in Rats

(A) Rats were divided into two groups (n = 5), one receiving ethanol (20%) and the other melatonin (7 μg) injected into the CA1 area. They were sacrificed 1 h later and the AICD SUMOylation level was determined. (B) Quantified results of (A) (t_1,8 = 6.7, p < 0.001). (C) Tissue lysates from the same animals were subjected to western blot analysis for the expression of PIAS1, NEP, and TTR. (D) Quantified results of (C) (t_1,8 = 8.12, p < 0.001 for PIAS1, t_1,8 = 9.77, p < 0.001 for NEP, and t_1,8 = 6.52, p < 0.001 for TTR). (E) Another batch of rats was divided into three groups (n = 5) and received DMSO (40%)+ethanol (20%), DMSO+melatonin (7 μg), or luzindole (1 μg)+melatonin (7 μg) injections to their CA1 area. They were sacrificed 1 h after the second injection and their CA1 tissue was subjected to western blot determination of PIAS1, NEP, and TTR expression. (F) Quantified results of (E) (F_2,12 = 38.2 for PIAS1, F_2,12 = 141.43 for NEP, and F_2,12 = 227.9 for TTR, all p < 0.001). (G) Rats were divided into three groups (n = 4) and received EGFP-vector transfection+ethanol (20%) injection, EGFP-vector transfection+melatonin injection (7 μg), and EGFP-AICDK43R transfection+melatonin injection (7 μg) into the CA1 area. The interval between these two injections was 24 h. They were sacrificed 1 h after melatonin (or ethanol) injection, and their CA1 tissue was subjected to western blot determination of NEP and TTR expression. (H) Quantified results of (G) (F_2,9 = 271.43, p < 0.001 for NEP and F_2,9 = 43.8, p < 0.001 for TTR). Data are expressed as individual values and mean ± SEM. ^#p < 0.001. EtOH, ethanol.