Fig 2: XAV939 reverses the effect of NGR1 in PC-12 cells. PC-12 cells were divided into four groups: control, LPS, LPS + NGR1, and LPS + NGR1 + XAV939. (a) MTT assay was applied to detect the viability of PC-12 cells in each group. (b and c) Flow cytometry analysis was used to assess the apoptosis of PC-12 cells in each group. (d) The protein levels of Bax, Bcl-2, and cleaved caspase-3 in PC-12 cells were assessed by western blotting. (e–g) The concentrations of IL-6, TNF-α, IL-1β in PC-12 cells were assessed by ELISA (Jh). The cytokine protein levels in PC-12 cells were measured by western blotting. **p < 0.01, ***p < 0.001.

Fig 3: Oral DSS administration caused colitis with GI tract-restricted inflammation in 10-week-old rats of both sexes.Length of the colon, defined as the end of the cecum to the anus, was measured as a macroscopic indicator of DSS-related colonic damage (A). Data are presented as box plots for n = 7–11 rats/group and were analyzed separately by sex using a two-sample t test (***p<0.001). Disease Activity Index (DAI) scores were also calculated daily (B). Average scores on each day for each sex and experimental group are depicted in the graph. Data for WT+DSS male and WT+DSS female were compared by two-way ANOVA with repeated measures (Day × Group, p<0.001). Individual t-tests were then used to compare groups at each treatment day (***p<0.001; ****p<0.0001); n = 7–11 rats/group. Furthermore, H&E-stained tissue sections from the descending colon of male WT rats were blindly scored in 3 randomly selected areas, and histology scores were generated (C). Data are presented as box plots for n = 10 rats/group and were analyzed by a two-sample t test (****p<0.0001). Serum IL-6 levels from male rats were also assessed by ELISA (D). No significant difference was noted between groups (by two-sample t test).

Fig 4: LINC00473 regulates IL-1β-stimulated C28/I2 apoptosis and the inflammatory response by regulating LY6E. (A) Overexpression efficacy of LY6E in C28/I2 cells was determined using reverse transcription-quantitative PCR. (B and C) Flow cytometry was performed to assess apoptosis in transfected C28/I2 cells. Protein expression levels of caspase-3, Bax and Bcl-2 in C28/I2 cells under the indicated transfections were (D) detected by western blotting and (E) semi-quantified. Concentrations of inflammatory cytokines (F) IL-6, (G) IL-8 and (H) TNF-α were measured using ELISAs in transfected C28/I2 cells. *P<0.05. LINC00473, long intergenic non-protein coding RNA 473; LY6E, lymphocyte antigen 6 locus E; si, small interfering RNA; NC, negative control; PI, propidium iodide

Fig 5: Levels of oxidative stress and inflammation in rats from control, model, 10 mg/kg eupafolin, 20 mg/kg eupafolin, 50 mg/kg eupafolin and nimodipine groups. The activities of (A) SOD, (B) MDA and (C) LDH in the brain tissues of rats at 24 h after cerebral I/R. The concentrations of (D) TNF-α, (E) IL-1β and (F) IL-6 in the serum of rats at 24 h after cerebral I/R. ***P<0.001 vs. control. #P<0.05, ##P<0.01 and ###P<0.001 vs. model. SOD, superoxide dismutase; MDA, malondialdehyde; LDH, lactate dehydrogenase; I/R, ischemia/reperfusion.