Fig 2: MAPK signaling pathway was activated under the infection of FMDV with reliance on viral replication. The protein expression of the marker proteins in MAPK signaling pathway after the infection of FMDV (A) or UV-treated FMDV (B) was confirmed by Western blot. The effects of MAPK/ERK signaling pathway inhibitor U0126-EtOH-EtOH on RNA levels (C), protein levels (D), and infectious FMDV virus titers (E) and the effects of MAPK/p38 signaling pathway inhibitor SB202190 on RNA levels (F), protein levels (G), and infectious FMDV virus titers (H). The intensity of pERK, p38, and p-JNK relative to GAPDH was measured by ImageLab software. GAPDH was served as the internal reference to normalize data. Three independent replicates were conducted for each sample. Data are expressed as the means ± SE (n = 3). *P < 0.01.

Fig 3: Validation of the effects of LR and GLN on LPS-induced inflammatory injury in cells (n = 3). (a) Changes in the relative TER values of Caco-2 cells for the completely differentiated monolayer; (b) FD4 content in lower compartments of Transwell plates. (c) Tight junction and apoptosis-related protein expression in Caco-2 cells. (d) p38, JNK and ERK1/2 phosphorylation in Caco-2 cells. (e) The tight junction and apoptosis-related protein expression when Caco-2 cells were treated with specific activators of p38, JNK and ERK1/2. ***: P < 0.001, **: 0.001 < P < 0.01, *: 0.01 < P < 0.05 indicate significance.

Fig 4: Effect of MG132 on apoptosis of NR8383 cells after H/R. (A) Apoptosis of NR8383 cells pretreated with the vehicle control (DMSO) or MG132 (10 µM) based on flow cytometry and (B) the quantified apoptotic rates. (C) Western blotting and (D) quantification of the protein levels of p-JNK and cleaved-caspase 3 in NR8383 cell extracts from the control group and MG132 group after H/R-2/2 and H/R-6/2. (All experiments were performed using the same protein extract, but due to the molecular weights of the target protein and the loading control protein being close, not all proteins were probed on the same membrane, but all experiments were performed as a single batch under identical conditions). *P<0.05, **P<0.01 and ***P<0.001. H/R, hypoxia/reoxygenation; H/R-2/2, hypoxia for 2 h then reoxygenation for 2 h; H/R-6/2, hypoxia for 6 h then reoxygenation for 2 h; p-, phosphorylated.

Fig 5: Potential targets of EYE-503 involved in retinal neurodegeneration. (a) The chemical structure of EYE-503. (b) Venn diagram showing the intersection target of EYE-503 and retinal neurodegeneration. (c) PPI network of the potential targets of EYE-503 during retinal neurodegeneration. (d) Bar plot diagram of GO enrichment analysis. (e) The top 10 items bubble plot of KEGG pathway analysis. (f) Western blot analysis of p38, p-p38, JNK, and p-JNK in retinal ganglion cells. GAPDH was used for normalization. (g) The phosphorylation levels of p38 and JNK in RGCs were evaluated by densitometric analysis. Relative p-p38 and p-JNK levels were calculated as the phosphorylated proteins levels divided by total proteins levels (n = 5, * p < 0.05 versus H2O2; one-way ANOVA).