Fig 2: Exosomes derived from M1 macrophages suppress GC progression. (A) Western blot analysis of the cell surface marker iNOS protein expression in LPS (100 ng/ml) + IFN-? (20 ng/ml)-induced macrophages (M1 macrophages) normalized to Na/K ATPase. (B) M1 macrophage membrane proteins quantified by image J and normalized with Na/K ATPase as well as statistics of iNOS gray value. (C) TEM observation of M1 macrophage-secreted exosomes (5,000×). (D) Dynamic light scattering detection of the diameter of exosomes. (E) Western blot analysis of the exosome surface markers (CD63 and HSP70) protein expression in M1 macrophages. (F) Flow cytometry of the exosome surface marker CD63 level. (G) Uptake of PKH76-labeled exosomes by GC cells was analyzed under a confocal fluorescence microscope (400×). (H) Tumor morphology in model mice with GC (n = 5). (I) Tumor volume in model mice with GC (n = 5). (J) Tumor weight in model mice with GC (n = 5). (K) Box plots of PDL1 expression in GC samples from TCGA analyzed by GEPIA (http://gepia.cancer-pku.cn/), wherein the left is GC tissues and the right is normal tissues, and the vertical axis is log2 [tags per million (TPM) + 1] (all PDL1 levels were normalized by TPM). (L) Western blot analysis of PD-L1 protein expression in model mice with GC normalized to GAPDH after protein quantitation by Image J (n = 5). *p < 0.05 compared with macrophages without the induction of IFN-? or M1 exosome or mouse GC models without any treatment; #p < 0.05 compared with mouse GC models treated with PBS. Measurement data was representative of three independently conducted experiments and expressed as the mean ± standard deviation. Comparisons between two groups are analyzed by non-paired t test, and comparisons among multiple groups are analyzed by one-way ANOVA. Post hoc test is conducted using Tukey’s test.

Fig 3: Influence of GAA on microglial phenotypic profiles and polarization. (A–C) qRT-PCR and Western blotting for expression levels of iNOS and CD86 in hippocampus tissues. (D–F) qRT-PCR and Western blotting for expression of Arg-1 and CD206 in hippocampus tissues. n=8. MCAO: stroke model; PSD: CUMS after MCAO; L-GAA: low dose (10 mg/kg) of GAA; M-GAA: median dose (20 mg/kg) of GAA; H-GAA: high dose (30 mg/kg) of GAA. **P<0.01 and ***P<0.001 compared with the sham group; &P<0.05 and &&P<0.01 compared with the MCAO group; #P<0.05 and ##P<0.01 compared with the PSD group.

Fig 4: Effect of SR-5 on NF?B/p65, iNOS, COX-2, and TNF-a protein expression in INDO/HCl-induced gastric mucosal injury mice. The Western blots are representative of NF?B/p65, iNOS, COX-2, and TNF-a proteins in mouse gastric tissue (A). The results are shown in the histogram of NF?B/p65 (B), iNOS (C), COX-2 (D), and TNF-a (E) proteins expressed as the ratio of the relative intensity of the level of expression of each protein to ß-actin. Each bar represents the mean ± SD for 7 mice. #Significant difference at p < .05 and ### at p < .001 compared to the control group (CTL). *Significant difference at p < .05, ** at p < .01, and *** at p < .001 compared to the INDO-induced gastric mucosal injury group.

Fig 5: Cardiac-specific overexpression of VCP increases iNOS expression and activity in TG mouse hearts compared to WT.(a) VCP TG mouse construct. (b) Representative immunoblots and protein level of VCP overexpression in mouse hearts, n = 8 per group. (c) mRNA level of iNOS in mouse hearts by qPCR, n = 5 per group. (d) Representative immunoblots and protein level of iNOS in mouse hearts, n = 8 per group. (e) iNOS activity in mouse heart tissues, n = 5 per group. (f) Protein level of eNOS in mouse hearts, n = 4 per group. GAPDH was used as loading control. *p < 0.05.**p < 0.01 vs. WT. Data are the mean ± SEM.