Fig 2: Effect of exosome uptake on macrophages.(A) Schematic showing mouse study with injection of either PBS, control-, or pIC-exosomes locally followed by excision of distal draining lymph nodes, isolation of macrophages and analysis of macrophage gene expression by next generation sequencing. (B) Relative expression of key M1 and housekeeping genes in macrophages after exposure to PBS, control- or pIC-exosomes. (C) Validation of key M1 genes in macrophages with control- or pIC-exosomes as compared to PBS by qRT-PCR. (D) Validation of M1 markers Nos2 and Il12 expression in lymph node sections after exposure to control- or pIC-exosomes. White scale bars, 50 µm, yellow scale bars, 10 µm.

Fig 3: Miconazole inhibits iNOS expression in in vivo and in vitro.The mRNA levels of iNOS were measured in mice hippocampus tissues (a) and 24-h LPS-treated (1 µg/mL) cultured astrocytes (c) and microglial BV2 cells (d) pretreated with MCZ (1.25, 2.5, 5, and 10 µM) for 1 h. iNOS proteins were detected by western blotting using specific antibodies in mice hippocampus tissues (b), cultured astrocytes (e), and microglial BV2 cells (f). ß-actin levels were measured to confirm equal protein loading. All experiments were performed three times with duplicate. The values on the western blot bands represent the arbitrary density measured by ImageJ. Data are expressed as the mean ± SEM (t-test, #p < 0.05 vs. Con, $p < 0.05 vs. MCZ, *p < 0.05 vs. MCZ + LPS). Significant difference from the control group (t-test, p < 0.05). #Significant difference from the LPS-treated group (p < 0.05). *Significant difference from the MCZ-treated group (p < 0.05).

Fig 4: TRPV4 activation by LPS triggers NO production in human bronchial EC. a Immunoblotting of 16HBE cells lysate probed for NOS isoforms. b Left, average intracellular Ca2+ responses of human bronchial epithelial cells to 20 µg ml-1 LPS and 10 nM GSK1016790A. The black, blue, and red solid traces correspond to data means of cells responding to LPS and GSK1016790A, GSK1016790A alone or to none of the stimuli, respectively. The thin dashed traces represent the means ± standard errors. Right, stack bar graph showing the percentages of cells responding to LPS and/or GSK1016790A (color-coded as the graph on the left; n = 116). c Immunoblotting of 16HBE cells lysate incubated in LPS (20 µg ml-1) or GSK1016790A (10 nM). Where indicated, cells were previously incubated with vehicle (Veh, 0.8% DMSO) or the TRPV4 inhibitor HC067047 (+HC, 10 µM). d Average intracellular Ca2+ signals (black solid traces) and normalized fluorescence of the NO-sensitive dye DAF-FM (F/F0, red solid traces) recorded in 16HBE cells (n = 52). The thin dashed traces represent the means ± standard errors. LPS, 20 µg ml-1; HC067047, 10 µM; GSK1016790A, 10 nM. e Average changes in intracellular Ca2+ (left panel) and NO production (right panel) induced by LPS. The dark cyan bars correspond to cells pre-incubated with the TRPV4 inhibitor HC067047 (10 µM). n > 74 per bar; **P < 0.01, two-tailed t-test. f Confocal microscopy images of 16HBE cells immunostained for iNOS. The arrow head points to an aggresome structure. Scale bar, 20 µm. g iNOS aggresomes were quantified using a custom-designed software. The graph shows the area of aggresomes relative to the total green-stained area determined from images recorded from preparations fixed in control condition or 5 min after application of 20 µg ml-1 LPS or 10 nM GSK1016790A. The panel on the right corresponds to cells pre-incubated with HC067047 (HC, 10 µM). Three randomly selected fields per condition were used for quantification. **P = 0.01, Dunn’s multiple comparison test

Fig 5: Chewing behavior during psychological stress improved oxidative stress in the tumor. (A) The expression level of iNOS protein in the tumor. (B) The expression level of 4HNE protein in the tumor. (C) The expression level of SOD2 protein in the tumor (** p < 0.01 vs. control group, ## p < 0.01 vs. stress group, n = 9/group). All data are expressed as mean ± SEM.