Fig 2: hLinfRNAs are expressed in a dose-dependent manner in THP1-macrophages (THP1-M?) under LPS induced inflammation. THP1-MF cells were treated with varying concentration of LPS (0.1- 1000 ng/mL, 4 h), total RNA was isolated and analyzed by RT-qPCR for expression of proinflammatory cytokines (IL6, IL-1ß) and top 5 hLinfRNAs. ß-Actin was used as loading control. Data represents mean ± SEM (n = 3).

Fig 3: (A) Representative Western blots showing the protein expression of IL-6 and Arg1 in BV2 cells cultured or not in the presence of AZE (1000 µM) for 6 or 24 h. (B,C) Violin plots of bands densities demonstrating the effects of AZE on the expression of IL-6 and Arg1 at the indicated times. Data reported is the mean ± SEM, from two independent experiments using separate batches of cells (n = 6). GAPDH was used as loading control. Ns = not significant. *** p < 0.001 or **** p < 0.0001 vs. Ctrl at the indicated time, as determined by ANOVA followed by Sidak’s post hoc test.

Fig 4: Effect of aging on cardiac IL-6 expression. (A) The expression of IL-6 in mice at 3 months, 15 months, 20 months, and 25 months of age was measured by Western blotting. N=5 in each group. * p<0.05 vs. the 3-month group. # p<0.05 vs. the previous group. (B) Plasma sIL-6R and sgp130 levels were detected in each group. N=5 in each group. * p<0.05 vs. the 3-month group. # p<0.05 vs. the previous group. (C) Effects of DOX treatment on IL-6 mRNA expression in WT Møs, lymphocytes, DCs, MCs, and CFs. N=5 in each group. * p<0.05 vs. the group. (D) The cardiac expression of both IL-6 and Mø markers in mice of different ages was detected in 25-month-old mice by immunofluorescence staining. N=5 in each group. (E) Double immunofluorescence staining for anti-F4/80 and anti-IL-6. N=5 in each group.

Fig 5: iPSC-EV alleviated IL-1ß-induced inflammation and cell death of chondrocytes. A. Representative western blot image showing the expression level of MMP13, IL-6, ADAMTS5, and collagen II in chondrocytes stimulated with IL-1ß and iPSC-EV. B.-E. Quantification of protein level from western blot analysis (n ?= ?3) F. Live and dead cell labelling observed using a fluorescence microscope, after incubating the chondrocyte with IL-1ß for 24 ?h. Scale bar ?= ?200 ?µm. G. Quantification of the percentage of apoptotic chondrocytes in live and dead cell assays (n ?= ?4) *Significant difference to control: *p ?< ?0.05, **p ?< ?0.01, ***p ?< ?0.005. #Significant difference between groups: ##p ?< ?0.01.Abbreviations: iPSC-EV: induced pluripotent stem cell-derived extracellular vesicles; IL-1ß: Interleukin 1 beta; MMP13: matrix metallopeptidase 13; IL-6: Interleukin 6; ADAMTS5: a disintegrin and metalloproteinase with thrombospondin motifs 5.