Exosomal miR-203 from bone marrow stem cells targets the SOCS3/NF-κB pathway to regulate neuroinflammation in temporal lobe epilepsy

Figure 4 MiR-203-3p regulates inflammation by targeting the suppression of cytokine signaling 3-mediated nuclear factor kappaB pathway. A: Luciferase reporter assay showing the relative firefly luciferase activity in HEK-293T cells co-transfected with wild-type or mutant suppression of cytokine signaling 3 3’ untranslated region reporter plasmids and miR-203-3p mimic or miR-203-3p; B: CCK8 assay to determine the IC₅₀ concentration of glutamate required to induce HT22 epilepsy; C: Quantitative polymerase chain reaction analysis of miR-203-3p expression in HT22 cells after treatment with miR-203-3p mimic or inhibitor; D: Quantitative polymerase chain reaction analysis of suppression of cytokine signaling 3 mRNA expression in HT22 cells under the conditions described in Figure 3C; E-L: Western blot analysis of key proteins involved in the nuclear factor kappaB pathway, including total and phosphorylated forms of inhibitor of kappa B kinase alpha/beta, nuclear factor kappaB inhibitor alpha, and p65, in HT22 cells treated with miR-203-3p mimics or inhibitors; M and N: Enzyme-linked immunosorbent assay measurements of the inflammatory cytokines interleukin-6 and tumor necrosis factor-α in the supernatant of HT22 cells treated as in Figure 3C. Data are presented as mean ± SEM of three independent experiments. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. SOCS3: Suppression of cytokine signaling 3; IKKα/β: Inhibitor of kappa B kinase alpha/beta; p-IKKα/β: Phospho-inhibitor of kappa B kinase alpha/beta; IKBα: Nuclear factor kappaB inhibitor alpha; p-IKBα: Phospho-nuclear factor kappaB inhibitor alpha; TNF-α: Tumor necrosis factor-α; IL-6: Interleukin-6.