Fig 2: The effects of perampanel (PER) and GYKI 52466 (GYKI) on Src family and its phosphorylations in chronic epilepsy rats. (A) Representative images for Western blot of Src family and its phosphorylation levels in the hippocampal tissues. (B–F) Quantifications of Src2 (B), p-Src Y416 (C), p-Src Y416/Src ratio (D), p-Src Y527 (E), and p-Src Y527/GRIA2 ratio (F) in the hippocampal tissues. Open circles indicate each individual value. Horizontal bars indicate mean value. Error bars indicate SEM (*, # p < 0.05 vs. control (Cont) and vehicle (Veh)-treated animals, respectively; one-way ANOVA with post hoc Bonferroni’s multiple comparison).

Fig 3: The effects of 3CAI co-treatment with perampanel (PER) and GYKI 52466 (GYKI) on surface expressions of GRIA1 and GRIA2 in chronic epilepsy rats. (A) Representative images for Western blot of total and membrane GRIA1 and GRIA2 expression levels. (B–H) Quantifications of total GRIA1 expression (B), membrane GRIA1 expression (C), membrane/total GRIA1 ratio (D), total GRIA2 expression (E), membrane GRIA2 expression (F), membrane/total GRIA2 ratio (G), and membrane GRIA1/GRIA2 ratio in the hippocampal tissues. Open circles indicate each individual value. Horizontal bars indicate mean value. Error bars indicate SEM (*, # p < 0.05 vs. control (Cont) and vehicle (Veh)-treated animals, respectively; one-way ANOVA with post hoc Bonferroni’s multiple comparison).

Fig 4: Effects of PDI knockdown on surface AMPAR expression in the hippocampus following KA injection. Control siRNA does not affect surface AMPAR expression in both saline- and KA-treated animals. PDI knockdown decreases surface expression of GluA1 and GluA2 without altering surface GluA1/GluA2 ratio in both saline- and KA-treated animals. PDI knockdown reduces surface AMPAR expression in KA-treated animals more than saline-treated animals. (a) Representative Western blot images for surface expressions of GluA1 and GluA2. (b–d) Quantitative analyses of the effects of PDI siRNA on surface GluA1 (b) and GluA2 (c) levels and surface GluA1/GluA2 ratio (d) following KA injection (*,#p < 0.05 vs. control siRNA vs. saline; n = 7, respectively; Kruskal–Wallis test followed by Tukey post-hoc test).

Fig 5: Effects of PDI knockdown on GluA2 phosphorylation and surface AMPAR expression in the hippocampus under physiological condition. As compared to control siRNA, PDI siRNA enhances GluA2 S880, but not Y869/873/876, phosphorylation in the mouse hippocampus. GluA2 S880 upregulations are observed in the dendrites and cell bodies of hippocampal neurons. However, PDI siRNA reduces surface expression of GluA1 and GluA2 without affecting surface GluA1/GluA2 ratio. (a) Representative Western blot images for GluA2, p-GluA2 Y869/873/876 and p-GluA2 S880 levels. (b–d) Quantitative analyses of the effects of PDI siRNA on GluA2 (b), p-GluA2 Y869/873/876 (c) and p-GluA2 S880 levels (d) based on the Western blot data (*p < 0.05 vs. control siRNA; n = 7, respectively; Mann–Whitney test). (e) Representative double immunostaining for NeuN and GluA2 S880 in CA1 and CA3 pyramidal neurons. (f) Representative Western blot images for surface expressions of GluA1 and GluA2. (g–i) Quantitative analyses of the effects of PDI siRNA on surface GluA1 (g) and GluA2 (h) levels and surface GluA1/GluA2 ratio (i) based on the Western blot data (*p < 0.05 vs. control siRNA; n = 7, respectively; Mann–Whitney test).