Fig 1: IL-1β, IL-6, TNF-α, L-glutamate levels, and neuronal apoptosis were decreased after miR-29c-3p mimic transfection in LPS-treated CTX-TNA2. (A) miR-29c-3p expression was measured via qRT-PCR after miR-29c-3p mimic transfection in LPS-treated CTX-TNA2 at 24 h. (B–E) IL-1β, IL-6, TNF-α, and L-glutamate levels were assessed using an assay kit according to the standard procedures. (F) Neurons cocultured with LPS-treated CTX-TNA2 (LPS group), LPS-treated CTX-TNA2 transfected NC mimic (NC mimic group), and LPS-treated CTX-TNA2 transfected miR-29c-3p mimic (miR-29c-3p mimic group), respectively. After cocultured at 48 h, the proliferation and apoptosis of neurons were evaluated via CCK8 and flow cytometry, respectively. *P < 0.05.
Fig 2: Proinflammatory cytokine profiles from the hippocampus (A, qRT-PCR) and serum (B, ELISA). At 8d post-SE, TNFα, IL-1β, and iNOS mRNAs were upregulated in the vehicle group, and SAR suppressed their levels. However, no significant differences were observed in their mRNA levels at 4 m post-SE. In serum, at 8d post-SE, TNFα, IL-1β, IL-6, and IL-12 were upregulated in the vehicle group, and SAR suppressed their levels. However, at 4 m, IL-1β and IL-6 levels were not altered in any groups but the increased TNFα and IL-12 levels were detected in the vehicle group with a significant reduction in the SAR group (B). n = 5–7; *#p < 0.05, **##p < 0.01, ***###p < 0.001, ****P < 0.0001; One-way ANOVA with Tukey’s post hoc test.
Fig 3: NFAT5 reverses the effect of XIST on LPS-treated CTX-TNA2. (A) NFAT5 expression were measured via western blot after transfection in LPS-treated CTX-TNA2 at 24 h. (B–E) IL-1β, IL-6, TNF-α, and L-glutamate levels were assessed using an assay kit according to the standard procedures after transfection in LPS-treated CTX-TNA2 at 24 h. (F, G) Neurons cocultured with LPS-treated CTX-TNA2 (LPS group), LPS-treated CTX-TNA2 transfected si-NC (LPS+si-NC group), and LPS-treated CTX-TNA2 transfected si-NFAT5 (LPS+si- NFAT5 group), LPS-treated si-XIST-CTX-TNA2 (LPS+si-XIST group), LPS-treated si-XIST-CTX-TNA2 transfected ov-NC (LPS+si-XIST+ov-NC group), and LPS-treated si-XIST-CTX-TNA2 transfected ov-NFAT5 (LPS+si-XIST+ ov-NFAT5 group), respectively. After cocultured at 48 h, the proliferation (F) and apoptosis (G) of neurons were evaluated via CCK8 and flow cytometry, respectively. *P < 0.05.
Fig 4: miR-29c-3p reverses the effect of XIST on LPS-treated CTX-TNA2. (A) XIST and miR-29c-3p expression were measured via qRT-PCR after si-XIST and miR-29c-3p mimic cotransfection in LPS-treated CTX-TNA2 at 24 h. (B–E) IL-1β, IL-6, TNF-α, and L-glutamate levels were assessed using an assay kit according to the standard procedures after si-XIST and miR-29c-3p mimic cotransfection in LPS-treated CTX-TNA2 at 24 h. (F) Neurons cocultured with LPS-treated si-XIST-CTX-TNA2 (LPS+si-XIST group), LPS-treated si-XIST-CTX-TNA2 transfected NC inhibitor (LPS+si-XIST+NC inhibitor group), and LPS-treated si-XIST-CTX-TNA2 transfected miR-29c-3p inhibitor (LPS+si-XIST+miR-29c-3p inhibitor group), respectively. After cocultured at 48 h, the proliferation and apoptosis of neurons were evaluated via CCK8 and flow cytometry, respectively. *P < 0.05.
Fig 5: IL-1β, IL-6, TNF-α, L-glutamate levels, and neuronal apoptosis were decreased after si-XIST transfection in LPS-treated CTX-TNA2. (A–D) IL-1β, IL-6, TNF-α, and L-glutamate levels were measured using ELISA kit according to the standard procedures. (E) Neurons cocultured with normal cultured CTX-TNA2 (control group), LPS-treated CTX-TNA2 (LPS group), LPS-treated CTX-TNA2 transfected si-NC (si-NC group), and LPS-treated CTX-TNA2 transfected si-XIST (si-XIST group), respectively. The proliferation and apoptosis of neurons was assessed via CCK8 and flow cytometry, respectively, after coculturing with LPS-treated si-XIST-transfected CTX-TNA2 for 48 h. *P < 0.05.
Supplier Page from Thermo Fisher Scientific for Rat IL-6 ELISA Kit