Fig. 7.

DNA methylation inhibitor treatment restores Afdn expression and BBB integrity by counteracting the hypermethylation of Afdn gene promoter induced by JEV supernatant. bEnd.3 cells were treated with mock or JEV supernatant for 48 h, followed by treatment with either DMSO or AZA for 24 h. (A) BSP method was used to detect changes in Afdn methylation level. (B) The changes in Afdn expression was measured by qPCR. (C) Western bolt was detected the expression of AFDN in protein levels. (D) Transwell experiment was conducted to assess the permeability of bEnd.3 cell. Data are represented as the mean values ± standard deviations (SD) of at least three biological replicates using Student’s t-test (*p < 0.05, **p < 0.01, ***p < 0.001, and ns: no significant) between the indicated groups